CDK2 inhibitors and uses thereof

Abstract

The present disclosure relates to compounds and pharmaceutical compositions, and methods for inhibiting the activity of CDK enzymes, in particular CDK2, with the compounds and compositions of the disclosure. The present disclosure further relates to, but is not limited to, methods for treating disorders associated with CDK2 signaling with the compounds and compositions of the disclosure.

Description

CDK2 INHIBITORS AND USES THEREOFCROSS-REFERENCE TO RELATED APPLICATIONS

[0001] This application claims priority to U.S. Provisional Application No. 63 / 547,220, filed November 3, 2023, the disclosure of which is herein incorporated by reference in its entirety.FIELD OF INVENTION

[0002] The present disclosure relates to compounds and pharmaceutical compositions capable of inhibiting the activity of CDK enzymes, in particular CDK2.BACKGROUND OF THE DISCLOSURE

[0003] Cyclin-dependent kinases (CDKs) are a family of serine / threonine kinases that are regulated by direct binding to cyclins. CDK levels remain relatively constant throughout the cell cycle, and it is the selective activation of specific CDKs that allows for the proper ordering of the steps in cell cycle progression. Activation of CDKs requires heterodimerization with regulatory subunits known as cyclins. Cell cycle deregulation is a common feature of human cancer.

[0004] Cyclin-dependent kinase 2 (CDK2) participates in a range of biological activities and is a key cell cycle regulator. For example, CDK2 is active from the late Gl-phase and throughout the S-phase, is involved in DNA damage response (DDR) through the homologous recombination (HR) pathway, and also regulates aspects of apoptotic pathways. Cyclin El (CCNE1), cyclin E2 (CCNE2), cyclin Al (CCNA1), and cyclin A2 (CCNA2), along with p21Cipl / Wafl, p27Kipl, and p57Kip2 (the cyclin dependent kinase inhibitors of the cyclin-CDK2 complex) are the main regulators of CDK2 activity. In cancer, persistent activation or hyperactivation of CDK2 can result from overexpression of the CDK2-binding cyclins El, E2, Al, or A2 or loss of activity of the cyclin-dependent kinase inhibitor proteins. (See S. Tadesse et al., Drug Discovery Today, Volume 25, Number 2 February 2020).

[0005] Amplification or overexpression of CCNE1 has been identified in ovarian and breast cancer (See Scaltriti, M. et al., Proc. Natl Acad. Sci. USA 108, 3761-3766 (2011) and Etemadmoghadam, D. et al. Proc. Natl Acad. Sci. USA 110, 19489-19494 (2013)). Poor outcomes in gastric, endometrial, and other cancers have been associated with overexpression or amplification of CCNE1 (See Ooi et al. Hum Pathol. (2017) 61 :58-67, and Noske et al, Oncotarget(2017) 8: 14794-14805). CCNE2 is frequently overexpressed in luminal breast cancers (Milioli et al. Endocrine Related Cancer (2020) 27: R93-R112).

[0006] Multiple pan-CDK inhibitors with activity against CDK2 and other CDKs have shown evidence of clinical activity, however they have also shown significant hematopoietic and gastrointestinal toxicities likely due to their inhibition of CDK1 and / or other CDK family members (Otto, T., and Sicinski, P., Nat. Review Cancer 2017; Kumar, K.S., et al. Blood 2015; Shapiro G.I., et al. Clin Cancer Research 2001).

[0007] While these findings indicate that CDK2 is a potential target for cancers with deregulated CDK2 activity, no agents selectively targeting CDK2 have been approved to date. Thus, there is a strong unmet need for developing new CDK2 inhibitors.SUMMARY OF THE DISCLOSURE

[0008] The present disclosure relates to compounds, pharmaceutical compositions, and methods for inhibiting the activity of CDK enzymes, in particular CDK2.

[0009] In one aspect, the present disclosure provides a compound having the structure selected from the group consisting ofa pharmaceutically acceptable salt thereof.

[0010] In some embodiments, the present disclosure provides a compound having the structure as shown in Tables 1-3, or a pharmaceutically acceptable salt thereof.

[0011] In another aspect, the present disclosure provides a pharmaceutical composition comprising the compound described in any of the above embodiments, or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier.

[0012] In another aspect, the present disclosure provides a pharmaceutical dosage form comprising the compound described in any of the above embodiments, or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition of the compound described in any of the above embodiments.

[0013] In yet another aspect, the present disclosure provides a method of targeting cyclin- dependent kinase 2 (CDK2) in a cell, the method comprising contacting the cell with an effective amount of the compound described in any of the above embodiments, or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition comprising the compound described in any of the above embodiments, or the pharmaceutical dosage form comprising the compound described in any of the above embodiments.

[0014] In some embodiments, the cell is a mammalian cell. In other embodiments, the cell is a tumor cell. In another embodiment, the tumor cell is a metastatic tumor cell.

[0015] In another aspect, the present disclosure provides a method of inhibiting CDK2 comprising contacting the CDK2 with an effective amount of the compound described in any of the above embodiments, or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition comprising the compound described in any of the above embodiments, or the pharmaceutical dosage form comprising the compound described in any of the above embodiments.

[0016] In some embodiments, the contacting is in a subject.

[0017] In yet another aspect, the present disclosure provides a method of treating a subject having, or at risk of developing, a disease or disorder associated with CDK2, comprising administering to the subject a therapeutically effective amount of the compound described in any of the above embodiments, or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition comprising the compound described in any of the above embodiments, or the pharmaceutical dosage form comprising the compound described in any of the above embodiments.

[0018] In yet another aspect, the present disclosure provides a method of treating a CDK2- mediated disorder or disease in a subject, the method comprising administering a therapeutically effective amount of the compound described in any of the above embodiments, or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition comprising the compound described in any of the above embodiments, or the pharmaceutical dosage form comprising the compound described in any of the above embodiments.

[0019] In some embodiments, the disorder is cancer. In some embodiments, the cancer treatable with a compound or composition of the present disclosure is a cancer of the lung, breast, gastrointestinal tract, esophagus, liver, biliary tract, bladder and other urinary tract tissues, pancreas, female and male genital organs, prostate, ovary, uterus, skin, lymphoid and blood cells, plasma cells (multiple myeloma) mesenchymal tissues (sarcomas), head and neck, visual system, brain and spinal cord, or melanoma. In some embodiments, the cancer can be selected from the group consisting of adrenal gland cancer, anal cancer, appendiceal cancer, ovarian cancer, uterine cancer, gastric cancer, breast cancer, lung cancer, bladder cancer, cervical cancer, stomach cancer, sarcoma cancer, liver cancer, esophageal cancer, laryngeal cancer, multiple myeloma, colorectal cancer, rectal cancer, skin cancer, pancreatic cancer, brain and spinal cord cancer, leukemia or lymphoma. In some embodiments, the compound is at least two-fold more selective for CDK2 over CDK1, CDK5, CDK6, CDK7, and / or CDK9.

[0020] In yet another aspect, the present disclosure provides a method of treating a cancer in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of the compound described in any of the above embodiments, or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition comprising the compound described in any of the above embodiments, or the pharmaceutical dosage form comprising the compound described in any of the above embodiments.

[0021] In some embodiments, the cancer can be characterized by deregulation of CDK2 or cyclin E.

[0022] In one embodiment, the cancer can be characterized by overexpression of cyclin El and / or cyclin E2. In some embodiments, the overexpression of cyclin El and / or cyclin E2 can be the overexpression of RNA and / or protein encoded by CCNE1 and / or CCNE2. In one embodiment, more than two copies of CCNE1 and / or CCNE2 can be present.

[0023] In some embodiments, the cancer can be characterized by a genomic alteration. In some embodiments, the genomic alteration can be characterized by one or more of the following: greater than the normal two copies (amplification) of the CCNE1 and / or CCNE2 genes, RBI loss of function mutation, FBXW7 loss of function mutation, MYC amplification, and / or KRAS mutation. The RBI loss of function can be attributed to function-disabling point mutation in the RB 1 gene, RB 1 gene deletion, and / or epigenetic repression of RB 1 gene expression (transcription). The FBXW7 loss of function can be attributed to function-disabling point mutation, gene deletion, and / or epigenetic repression of FBXW7 gene expression (transcription). MYC gene amplification or protein stabilization can lead to MYC overexpression. The MYC overexpression can be attributed to one or more of the MYC family members, c-MYC, L-MYC and N-MYC.

[0024] In some embodiments, the cancer can be metastatic cancer. In one embodiment, the cancer can be characterized by its origination in or metastasis to the brain. In one embodiment, the cancer is a cancer of the lung, breast, esophagus, and melanoma that has metastasized to the brain.

[0025] In some embodiments, the cancer can be selected from the group consisting of adrenal gland cancer, anal cancer, appendiceal cancer, ovarian cancer, uterine cancer, gastric cancer, breast cancer, lung cancer, bladder cancer, cervical cancer, stomach cancer, sarcoma cancer, liver cancer, esophageal cancer, laryngeal cancer, multiple myeloma, colorectal cancer, rectal cancer, skin cancer, pancreatic cancer, brain or spinal cord cancer, leukemia or lymphoma.

[0026] In some embodiments, the cancer is platinum -resistant and / or platinum-refractory. In some embodiments, the cancer has progressed despite platinum treatment.

[0027] In various embodiments, the adrenal gland cancer can be adrenocortical carcinoma or pheochromocytoma. The breast cancer can be hormone receptor (HR)-positive / HER2-negative breast cancer; HR-positive / human epidermal growth factor receptor 2 (HER2)-positive breast cancer; triple negative breast cancer (TNBC), or inflammatory breast cancer. The breast cancer can be drug-naive breast cancer, endocrine resistant breast cancer, trastuzumab resistant breastcancer, breast cancer demonstrating primary or acquired resistance to CDK4 / CDK6 inhibition, platinum-resistant breast cancer or platinum-refractory breast cancer. The breast cancer can be advanced or metastatic breast cancer. The liver cancer can be hepatocellular carcinoma. The lung cancer can be small cell lung cancer, non-small cell lung cancer, or large cell lung cancer. The non-small cell lung cancer can be squamous cell carcinoma or adenocarcinoma. The laryngeal cancer can be laryngeal squamous cell carcinoma. The skin cancer can be melanoma, basal cell carcinoma, squamous cell carcinoma, or Merkel cell cancer. The uterine cancer can be endometrial cancer or uterine sarcoma. In some embodiments, the uterine cancer (e g., endometrial cancer) is platinum -resistant or platinum-refractory endometrial cancer. In some embodiments, the ovarian cancer is platinum-resistant or platinum-refractory ovarian cancer. The brain cancer can be selected from glioma, astrocytoma, meningioma, glioblastoma multiforme, medulloblastoma, ependymoma, oligodendroglioma, craniopharyngioma, pituitary adenoma, Schwannoma, anaplastic astrocytoma, germ cell tumor, primitive neuroectodermal tumor, chordoma, haemangioblastoma, optic nerve glioma, subependymoma, and germinoma.

[0028] In various embodiments, the subject can be a human. The subject can be an adult. The subject can be a pediatric subject 0 to 18 years of age.

[0029] In some embodiments, the compound, the pharmaceutical composition, or the pharmaceutical dosage form can be administered orally, intravenously, or subcutaneously.

[0030] In some embodiments of any of the above methods, the method can further comprise administering an effective amount of a second therapeutic agent. The second therapeutic agent can be an anti -neoplastic agent. The anti -neoplastic agent can be selected from the group consisting of an antibody, an antibody-drug conjugate, a DNA synthesis inhibitor, a DNA- intercalating agent, a platinum-based agent, topoisomerase I inhibitor, topoisomerase II inhibitor, an alkylating agent, an anti-microtubule agent, an anti-mitotic agent, a taxane-r elated anti- neoplastic agent, an anti-metabolite, an anti-tumor plant alkaloid, an anti-estrogen agent, a protein kinase inhibitor, a phosphatidylinositol 3-kinase inhibitor, an immunomodulator, a histone deacetylase inhibitor, a KRAS inhibitor, an immunotherapeutic agent, or an epigenetic modulator. The antibody-drug conjugate can comprise sacituzumab govitecan (Trodelvy), mirvetuximab soravtansine (Elahere), trastuzumab deruxtecan (Enhertu), enfortumab vedotin (Padcev), or tisotumab vedotin (Tivdak). The DNA synthesis inhibitor can comprise capecitabine, gemcitabine, nelarabine, or hydroxycarbamide. The platinum-based agent can comprise carboplatin, oxaliplatinor cisplatin. The topoisomerase I inhibitor can comprise camptothecin, irinotecan or topotecan. The DNA-intercalating agent can comprise doxorubicin or liposomal doxorubicin (Doxil). The topoisomerase II inhibitor can comprise etoposide or teniposide. The alkylating agent can comprise temozolomide, dactinomycin, melphalan, altretamine, carmustine, bendamustine, busulfan, lomustine, chlorambucil, cyclophosphamide, dacarbazine, altretamine, ifosfamide, or procarbazine. The anti -microtubule agent can comprise estramustine. The taxane anti-neoplastic agent can comprise paclitaxel, docetaxel, cabazitaxel larotaxel, or abraxane. The anti-metabolite can comprise 5-fluorouracil, 6-thioguanine, pemetrexed, cytarabine, hydroxyurea, fludarabine, floxuridine, cladribine, pentostatin, or methotrexate. The anti-tumor plant alkaloid can comprise vinblastine, vincristine, or vinorelbine. The anti-estrogen agent can comprise tamoxifen, toremifene, raloxifene, lasofoxifene, afimoxifene, arzoxifene, bazedoxifene, fispemifene, ormeloxifene, ospemifene, tesmilifene, trilostane, CHF 4227, fulvestrant, elacestrant, giredestrant, amcenestrant, camizestrant, ataraestane, formestane, exemestane, letrozole, anastrozole, fadrozole; gonadotropin-releasing hormone, leuprolide, leuprolide acetate, enzalutamide, abiraterone acetate, or bicalutamide. The protein kinase inhibitor can comprise a CDK4 / 6 inhibitor (e g., palbociclib, ribociclib, trilaciclib, or abemaciclib), alvocidib, crizotinib, NVP-TAE684, ceritinib, alectinib, brigatinib, entrectinib, lorlatinib, vemurafenib, dabrafenib, infigratinib, dovitinib, erdafitinib, BLU-554, AZD4547, trametinib, cobimetinib, binimetinib, selumetinib, ulixertinib, MK-8353, LY-3214966, bevacizumab, axitinib, aflibercept, brivanib, motesanib, pasireotide, sorafenib, erlotinib, linifanib, sunitinib, pazopanib, gefitinib, osimertinib, cetuximab, panitumumab, trastuzumab, neratinib, lapatinib, or cabozantinib. The histone deacetylase inhibitor can comprise voninostat. The phosphatidylinositol 3-kinase inhibitor can comprise apelisib (Piqray), duvelisib (Copiktra), copanlisib (Aliqopa), or idelalisib (Zydelig). The KRAS inhibitor can comprise AMG510, MRTX849, JNJ-74699157 / ARS- 3248, Bl 1701963, Bl 1823911, BAY- 293, GDC-6036, MRTX1133, a RAS(ON) inhibitor, or combinations thereof. The immunotherapeutic agent can target at least one of PD1, PD-L1, PD-L2, CTLA-4, LAG3, TIM3, VISTA, BTLA, TIGIT, LAIR1, CD 160, 2B4, 4- IBB, GITR, 0X40, or TGF beta receptor. In some embodiments, the immunotherapeutic agent comprises a CTLA-4 inhibitor, a PD1 inhibitor, a PD- L1 inhibitor, a LAG3 inhibitor, or a 4- IBB agonist. The CTLA-4 inhibitor can comprise tremelimumab or ipilimumab. The PD1 inhibitor can comprise nivolumab or pembrolizumab. The PDL1 inhibitor can comprise atezolizumab, avelumab, or durvalumab. The LAG3 inhibitor cancomprise BMS-986016. In some embodiments, the immunotherapeutic agent comprises a chimeric antigen receptor (CAR) T-cell therapy. The epigenetic modulator may comprise EZH2a, ARID 1 A, ARID2, PRMT1-9, BRD4, IDH1 / 2, or BCL6.

[0031] These and other aspects of the present disclosure will become apparent to those skilled in the art after a reading of the following detailed description, including the appended claims.DETAILED DESCRIPTION

[0032] Detailed embodiments of the present disclosure are disclosed herein; however, it is to be understood that the disclosed embodiments are merely illustrative of the disclosure that may be embodied in various forms. In addition, each of the examples given in connection with the various embodiments of the disclosure is intended to be illustrative, and not restrictive. Therefore, specific structural and functional details disclosed herein are not to be interpreted as limiting, but merely as a representative basis for teaching one skilled in the art to variously employ the present disclosure.

[0033] Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this disclosure belongs. For purposes of interpreting this specification, the following description of terms will apply and whenever appropriate, terms used in the singular will also include the plural and vice versa. All patents, applications, published applications and other publications are incorporated by reference in their entirety. In the event that any description of terms set forth conflicts with any document incorporated herein by reference, the description of term set forth below shall control.Definitions

[0034] As used in this specification and the appended claims, the singular forms “a”, “an”, and “the” include plural references unless the context clearly dictates otherwise. Thus, for example, a reference to “a method” includes one or more methods, and / or steps of the type described herein and / or which will become apparent to those persons skilled in the art upon reading this disclosure.

[0035] As used herein, the term "about," when used in reference to a particular recited numerical value, means that the value may vary from the recited value by no more than 10%. Forexample, as used herein, the expression "about 100" includes 90 and 1 10 and all values in between (e g., 91, 92, 93, 94, 95, 96, 97, 98, 99, etc.).

[0036] Herein, the use of terms such as “having,” “has,” “including,” or “includes” are open-ended and are intended to have the same meaning as terms such as “comprising” or “comprises” and not preclude the presence of other structure, material, or acts. Similarly, though the use of terms such as “can” or “may” are intended to be open-ended and to reflect that structure, material, or acts are not necessary, the failure to use such terms is not intended to reflect that structure, material, or acts are essential. To the extent that structure, material, or acts are presently considered to be essential, they are identified as such.

[0037] By “comprising” or “containing” or “including” is meant that at least the named compound, element, particle, or method step is present in the composition or article or method, but does not exclude the presence of other compounds, materials, particles, method steps, even if the other such compounds, material, particles, method steps have the same function as what is named.

[0038] It is also to be understood that the mention of one or more method steps does not preclude the presence of additional method steps or intervening method steps between those steps expressly identified.

[0039] For purposes of this disclosure, the chemical elements are identified in accordance with the Periodic Table of the Elements, CAS version, Handbook of Chemistry and Physics, 75th Ed. Additionally, general principles of organic chemistry are described in “Organic Chemistry”, Thomas Sorrell, University Science Books, Sausalito: 1999, and “March’s Advanced Organic Chemistry”, 5th Ed., Ed.: Smith, M.B. and March, J., John Wiley & Sons, New York: 2001, the entire contents of which are hereby incorporated by reference. Chemical names, common names, and chemical structures may be used interchangeably to describe the same structure. If a chemical compound is referred to using both a chemical structure and a chemical name, and an ambiguity exists between the structure and the name, the structure predominates.

[0040] Compounds described herein may contain one or more asymmetric centers and may thus give rise to enantiomers, diastereomers, and other stereoisomeric forms. Each chiral center may be defined, in terms of absolute stereochemistry, as (R)- or (S)-. This technology is meant to include all such possible isomers, as well as mixtures thereof, including racemic and optically pure forms. Optically active (R)- and (S)-, (-)- and (+)-, or (D)- and (L)- isomers may be prepared using chiral synthons or chiral reagents, or resolved using conventional techniques. When thecompounds described herein contain olefinic double bonds or other centers of geometric asymmetry, and unless specified otherwise, it is intended that the compounds include both E and Z geometric isomers. Likewise, all tautomeric forms are also intended to be included.

[0041] The term “tautomer” as used herein refers to one of two or more structural isomers which exist in equilibrium and which are readily converted from one isomeric form to another. It is understood that tautomers encompass valence tautomers and proton tautomers (also known as prototropic tautomers). Valence tautomers include interconversions by reorganization of some of the bonding electrons. Proton tautomers include interconversions via migration of a proton, such as keto-enol and imine-enamine isomerizations. Unless otherwise stated, all tautomers of the compounds described herein are within the scope of the disclosure.

[0042] The term “isotopic substitution” as used herein refers to the substitution of an atom with its isotope. The term “isotope” as used herein refers to an atom having the same atomic number as that of atoms dominant in nature but having a mass number (neutron number) different from the mass number of the atoms dominant in nature. It is understood that a compound with an isotopic substitution refers to a compound in which at least one atom contained therein is substituted with its isotope. Atoms that can be substituted with its isotope include, but are not limited to, hydrogen, carbon, and oxygen. Examples of the isotope of a hydrogen atom include2H (Deuterium, also represented as D) and3H (Tritium, also represented as T). Examples of the isotope of a carbon atom include13C and14C. Examples of the isotope of an oxygen atom include18O. Unless otherwise stated, all isotopic substitution of the compounds described herein are within the scope of the disclosure. Such compounds are useful, for example, as analytical tools, as probes in biological assays, or as therapeutic agents in accordance with the present disclosure.

[0043] As used herein, "crystalline form" is meant to refer to a certain lattice configuration of a crystalline substance. Different crystalline forms (polymorphs) of the same substance typically have different crystalline lattices (e.g., unit cells) which are attributed to different physical properties that are characteristic of each of the crystalline forms. In some instances, different lattice configurations have different water or solvent content. Crystalline forms of a substance include both solvated (e.g., hydrated) and non-solvated (e.g., anhydrous) forms. A hydrated form is a crystalline form that includes water in the crystalline lattice. Hydrated forms can be stoichiometric hydrates, where the water is present in the lattice in a certain water / molecule ratio such as forhemihydrates, monohydrates, dihydrates, etc. Hydrated forms can also be non-stoichiometric, where the water content is variable and dependent on external conditions such as humidity.

[0044] The different crystalline lattices can be identified by solid state characterization methods such as by X-ray powder or thin film diffraction (XRD). Other characterization methods such as Raman spectroscopy, differential scanning calorimetry (DSC), thermogravimetric analysis (TGA), dynamic vapor sorption (DVS), solid state NMR, and the like further help identify the crystalline form as well as help determine stability and solvent / water content. Typically, different crystalline forms of the same substance have different bulk properties relating to, for example, hygroscopicity, solubility, stability, and the like. Forms with high melting points often have good thermodynamic stability which is advantageous in prolonging shelf-life of formulations containing the solid form. Forms with lower melting points often are less thermodynamically stable, but are advantageous in that they have increased water solubility, translating to increased drug bioavailability. Forms that are weakly or non-hygroscopic are desirable for their stability to heat and humidity and are resistant to degradation during long storage. Anhydrous forms are often desirable because they can be consistently made without concern for variation in weight or composition due to varying solvent or water content. On the other hand, hydrated or solvated forms can be advantageous in that they are less likely to be hygroscopic and may show improved stability to humidity under storage conditions. With respect to XRD, the term "peak" refers to a reflection having a relative height / intensity of at least about 4% of the maximum peak height / intensity. Moreover, instrument variation and other factors can affect the 2-theta values. Thus, peak assignments, such as those reported herein, can vary by plus or minus about 0.2° (2-theta), and the term "substantially" as used in the context of XRD herein is meant to encompass the above- mentioned variations.

[0045] When a compound recrystallizes from a solution, slurry, within a porous media, or other means, the compound may crystallize with different spatial lattice arrangements, a property referred to as “polymorphism.” The different crystal forms are individually referred to as a “polymorph” or “form.” Different polymorphic forms of a compound may differ from each other with respect to one or more physical property, such as solubility, true density, crystal shape, compaction behavior, flow properties, lethality, solid state stability, and the like.

[0046] As used herein, the term “pharmaceutically acceptable salt” refers to those salts which are, within the scope of sound medical judgment, suitable for use in contact with the tissuesof humans and lower animals without undue toxicity, irritation, allergic response and the like, and are commensurate with a reasonable benefit / risk ratio. Exemplary pharmaceutically acceptable salts are found, e.g., in Berge, et al. (J. Pharm. Sci. 1977, 66(1), 1; and Gould, P.L., Int. J. Pharmaceutics 1986, 33, 201-217; (each hereby incorporated by reference in its entirety).

[0047] Pharmaceutically acceptable salts of the compounds described herein include those derived from suitable inorganic and organic acids and bases. Examples of pharmaceutically acceptable, nontoxic acid addition salts are salts of an amino group formed with inorganic acids such as hydrochloric acid, hydrobromic acid, phosphoric acid, sulfuric acid and perchloric acid or with organic acids such as acetic acid, oxalic acid, maleic acid, tartaric acid, citric acid, succinic acid or malonic acid or by using other methods used in the art such as ion exchange. Other pharmaceutically acceptable salts include adipate, alginate, ascorbate, aspartate, benzenesulfonate, benzoate, bisulfate, borate, butyrate, camphorate, camphorsulfonate, citrate, cyclopentanepropionate, digluconate, dodecyl sulfate, ethanesulfonate, formate, fumarate, glucoheptonate, glycerophosphate, gluconate, hemisulfate, heptanoate, hexanoate, hydroiodide, 2- hydroxy-ethanesulfonate, lactobionate, lactate, laurate, lauryl sulfate, malate, maleate, malonate, methanesulfonate, 2-naphthalenesulfonate, nicotinate, nitrate, oleate, oxalate, palmitate, pamoate, pectinate, persulfate, 3 -phenyl propionate, phosphate, pivalate, propionate, stearate, succinate, sulfate, tartrate, thiocyanate, / >-toluenesulfonate, undecanoate, valerate salts, and the like.

[0048] Salts derived from appropriate bases include alkali metal, alkaline earth metal, ammonium and N+(Ci-4alkyl)4 salts. Representative alkali or alkaline earth metal salts include sodium, lithium, potassium, calcium, magnesium, and the like. Further pharmaceutically acceptable salts include, when appropriate, nontoxic ammonium, quaternary ammonium, and amine cations formed using counterions such as halide, hydroxide, carboxylate, sulfate, phosphate, nitrate, loweralkyl sulfonate and aryl sulfonate.

[0049] Pharmaceutically acceptable salts are also intended to encompass hemi-salts, wherein the ratio of compound:acid is respectively 2: 1. Exemplary hemi-salts are those salts derived from acids comprising two carboxylic acid groups, such as malic acid, fumaric acid, maleic acid, succinic acid, tartaric acid, glutaric acid, oxalic acid, adipic acid and citric acid. Other exemplary hemi-salts are those salts derived from diprotic mineral acids such as sulfuric acid. Exemplary hemi-salts include, but are not limited to, hemimaleate, hemifumarate, and hemisuccinate.

[0050] An “effective amount”, “sufficient amount” or “therapeutically effective amount” as used herein is an amount of a compound that is sufficient, when administered to a subject or population suffering from or susceptible to a disease, disorder, and / or condition in accordance with a therapeutic dosing regimen, to treat (e.g., effect beneficial or desired results, including clinical results) the disease, disorder, and / or condition. As such, the effective amount may be sufficient, e.g., to reduce or ameliorate the severity and / or duration of afflictions related to CDK2 signaling, or one or more symptoms thereof, prevent the advancement of conditions or symptoms related to afflictions related to CDK2 signaling, or enhance or otherwise improve the prophylactic or therapeutic effect(s) of another therapy. An effective amount also includes the amount of the compound that avoids or substantially attenuates undesirable side effects.

[0051] The terms “enhance” or “promote,” or “increase,” or “expand,” or “improve” refer generally to the ability of a composition contemplated herein to produce, elicit, or cause a greater physiological response (i.e., downstream effects) compared to the response caused by either vehicle or a control molecule / composition. A measurable physiological response may include an increase in immune cell expansion, activation, effector function, persistence, and / or an increase in tumor cell death killing ability, among others apparent from the understanding in the art and the description herein. In certain embodiments, an “increased” or “enhanced” amount can be a “statistically significant” amount, and may include an increase that is 1.1, 1.2, 1.5, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 30 or more times (e.g., 500, 1000 times) (including all integers and decimal points in between and above 1, e.g., 1.5, 1.6, 1.7. 1.8, etc.) the response produced by vehicle or a control composition.

[0052] The terms “decrease” or “lower,” or “lessen,” or “reduce,” or “abate”, or “attenuate”, or “suppress” refer generally to the ability of composition contemplated herein to produce, elicit, or cause a lesser physiological response (i.e., downstream effects) compared to the response caused by either vehicle or a control molecule / composition. In certain embodiments, a “decrease” or “reduced” amount can be a “statistically significant” amount, and may include a decrease that is 1.1, 1.2, 1.5, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 30 or more times (e.g., 500, 1000 times) (including all integers and decimal points in between and above 1, e.g., 1.5, 1.6, 1.7. 1.8, etc.) the response (reference response) produced by vehicle or a control composition.

[0053] The terms “treat” or “treatment” of a state, disorder or condition include: (1) preventing, delaying, or reducing the incidence and / or likelihood of the appearance of at least oneclinical or sub-clinical symptom of the state, disorder or condition developing in a subject that may be afflicted with or predisposed to the state, disorder or condition, but does not yet experience or display clinical or subclinical symptoms of the state, disorder or condition; or (2) inhibiting the state, disorder or condition, i.e., arresting, reducing or delaying the development of the disease or a relapse thereof or at least one clinical or sub-clinical symptom thereof; or (3) relieving the disease, i.e., causing regression of the state, disorder or condition or at least one of its clinical or sub-clinical symptoms. The benefit to a subject to be treated is either statistically significant or at least perceptible to the patient or to the physician.

[0054] The terms “effective amount” or “therapeutically effective amount” refer to a quantity and / or concentration of a composition containing an active ingredient (e.g., CDK2 inhibitor compound) that when administered into a patient either alone (i.e., as a monotherapy) or in combination with additional therapeutic agents, yields a significant decrease in disease progression, for example, by ameliorating or eliminating symptoms and / or the cause of the disease. An effective amount may be an amount that relieves, lessens, or alleviates at least one symptom or biological response or effect associated with a disease or disorder, prevents progression of the disease or disorder, or improves physical functioning of the patient. A therapeutically effective amount of a composition containing an active agent may vary according to factors such as the disease state, age, sex, and weight of the individual, and the ability of the active agent to elicit a desired response in the individual. A therapeutically effective amount is also one in which any toxic or detrimental effects of the active agent are outweighed by the therapeutically beneficial effects. A therapeutically effective amount may be delivered in one or more administrations. A therapeutically effective amount refers to an amount effective, at dosages and for periods of time necessary, to achieve the desired therapeutic and / or prophylactic result.

[0055] The terms “individual”, “subject” and “patient” are used interchangeably herein to refer to an animal; for example a mammal. The terms include human and veterinary subjects. In some embodiments, methods of treating mammals, including, but not limited to, humans, rodents, simians, felines, canines, equines, bovines, porcines, ovines, caprines, mammalian laboratory animals, mammalian farm animals, mammalian sport animals, and mammalian pets, are provided. The subject can be male or female and can be any suitable age, including infant, juvenile, adolescent, adult, and geriatric subjects. In some embodiments, a subject can be a subject in need of treatment for a disease or disorder. In particular embodiments, the subject is a human.

[0056] As used herein, the human subject can be an adult, older adult (e g., geriatric), or a pediatric subject. “Adult” subjects mean 21 years to less than 65 years and “older adult” or "geriatric" subjects range from 65 years to less than 74 years; 75 years to less than 84 years; and greater than 85 years. “Pediatric” subjects include neonates (i.e., from birth through the first 28 days of life), infants (29 days to less than 2 years), children (2 years to less than 12 years), or adolescents (aged 12 years through 21 years).

[0057] The phrase “in need thereof’ refers to the need for symptomatic or asymptomatic relief from conditions related to CDK2 signaling activity or that may otherwise be relieved by the compounds and / or compositions of the disclosure.Compounds of the Disclosure

[0058] In some embodiments, the compound can have a structure selected from the group consisting of.0.a pharmaceutically acceptable salt thereof.

[0059] In some exemplary, non-limiting embodiments, the compound of the disclosure can be selected from a compound of Tables 1-3.

[0060] In some embodiments, the compounds of the disclosure are CDK2 inhibitors. In some embodiments, the compounds of the disclosure are selective CDK2 inhibitors. As used herein, the term “selective CDK2 inhibitor” means a compound which selectively inhibits CDK2 over other CDKs and the kinome. Said another way, a selective CDK2 inhibitor has no activity or lower activity against other CDKs and the kinome. A selective CDK2 inhibitor’s inhibitory activity against CDK2 is more potent in terms of ICso value (i.e., the ICso value is lower) when compared with its inhibitory activity against other CDKs and many other kinases. Potency can be measured using known biochemical assays.

[0061] In some embodiments, the activity of a compound described herein (e g., the compounds of Tables 1-3) as an inhibitor of a CDK kinase, for example, CDK2, or a mutant thereof, may be assayed in vitro, in vivo or in a cell line. In vitro assays include assays that determine inhibition of either the phosphorylation activity and / or the subsequent functional consequences, or ATPase activity of activated CDK2, or a mutant thereof. Alternative in vitro assays quantitate the ability of the inhibitor to bind to CDK2. Inhibitor binding may be measured by radiolabeling the inhibitor prior to binding, isolating the inhibitor / CDK2 complex and determining the amount of radiolabel bound. Alternatively, inhibitor binding may be determined by running a competition experiment where new inhibitors are incubated with CDK2 bound to known radioligands. Representative in vitro and in vivo assays useful in assaying a CDK2 inhibitor include those described and disclosed in the patent and scientific publications described herein. Detailed conditions for assaying a compound described herein as an inhibitor of CDK2, or a mutant thereof, are set forth in the Examples below.

[0062] In some embodiments, the compounds of the disclosure are selective against CDK2 versus CDK1. In some embodiments, compounds show at least 2-fold selectivity for CDK2 versusCDK1 In some embodiments, compounds show at least 5-fold selectivity for CDK2 versus CDK1 . In some such embodiments, compounds show at least 10-fold selectivity for CDK2 versus CDK1. In other embodiments, compounds show at least 20-fold selectivity for CDK2 versus CDK1. In some embodiments, compounds show at least 30-fold selectivity for CDK2 versus CDK1. In certain embodiments, compounds show at least 40-fold selectivity for CDK2 versus CDK1. In other embodiments, compounds show at least 50-fold selectivity for CDK2 versus CDK1. For example, compounds show at least 100-fold selectivity for CDK2 versus CDK1.

[0063] In some embodiments, the compounds of the disclosure are selective against CDK2 versus CDK5. In some embodiments, compounds show at least 2-fold selectivity for CDK2 versus CDK5. In some embodiments, compounds show at least 5-fold selectivity for CDK2 versus CDK5. In some such embodiments, compounds show at least 10-fold selectivity for CDK2 versus CDK5. In other embodiments, compounds show at least 20-fold selectivity for CDK2 versus CDK5. In some embodiments, compounds show at least 30-fold selectivity for CDK2 versus CDK5. In certain embodiments, compounds show at least 40-fold selectivity for CDK2 versus CDK5. In other embodiments, compounds show at least 50-fold selectivity for CDK2 versus CDK5. For example, compounds show at least 100-fold selectivity for CDK2 versus CDK5.

[0064] In some embodiments, the compounds of the disclosure are selective against CDK2 versus CDK6. In some embodiments, compounds show at least 2-fold selectivity for CDK2 versus CDK6. In some embodiments, compounds show at least 5-fold selectivity for CDK2 versus CDK6. In some such embodiments, compounds show at least 10-fold selectivity for CDK2 versus CDK6. In other embodiments, compounds show at least 20-fold selectivity for CDK2 versus CDK6. In some embodiments, compounds show at least 30-fold selectivity for CDK2 versus CDK6. In certain embodiments, compounds show at least 40-fold selectivity for CDK2 versus CDK6. In other embodiments, compounds show at least 50-fold selectivity for CDK2 versus CDK6. For example, compounds show at least 100-fold selectivity for CDK2 versus CDK6.

[0065] In some embodiments, the compounds of the disclosure are selective against CDK2 versus CDK7. In some embodiments, compounds show at least 2-fold selectivity for CDK2 versus CDK7. In some embodiments, compounds show at least 5-fold selectivity for CDK2 versus CDK7. In some such embodiments, compounds show at least 10-fold selectivity for CDK2 versus CDK7. In other embodiments, compounds show at least 20-fold selectivity for CDK2 versus CDK7. In some embodiments, compounds show at least 30-fold selectivity for CDK2 versus CDK7. Incertain embodiments, compounds show at least 40-fold selectivity for CDK2 versus CDK7. In other embodiments, compounds show at least 50-fold selectivity for CDK2 versus CDK7. For example, compounds show at least 100-fold selectivity for CDK2 versus CDK7.

[0066] In some embodiments, the compounds of the disclosure are selective against CDK2 versus CDK9. In some embodiments, compounds show at least 2-fold selectivity for CDK2 versus CDK9. In some embodiments, compounds show at least 5-fold selectivity for CDK2 versus CDK9. In some such embodiments, compounds show at least 10-fold selectivity for CDK2 versus CDK9. In other embodiments, compounds show at least 20-fold selectivity for CDK2 versus CDK9. In some embodiments, compounds show at least 30-fold selectivity for CDK2 versus CDK9. in certain embodiments, compounds show at least 40-fold selectivity for CDK2 versus CDK9. In other embodiments, compounds show at least 50-fold selectivity for CDK2 versus CDK9. For example, compounds show at least 100-fold selectivity for CDK2 versus CDK9.Methods of Use

[0067] In some embodiments, the CDK2 inhibitors of the present disclosure (e.g., the compounds of Tables 1-3) are used in a method of targeting cyclin-dependent kinase 2 (CDK2) in a cell. The method can include contacting the cell with an effective amount of a compound described herein, or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition of a compound described herein, or the pharmaceutical dosage form of a compound described herein.

[0068] In certain embodiments, the CDK2 inhibitors of the present disclosure (e g., the compounds of Tables 1-3) are used in a method of inhibiting cyclin-dependent kinase 2 (CDK2) in a cell. The method can include contacting the cell with an effective amount of a compound described herein, or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition of a compound described herein, or the pharmaceutical dosage form of a compound described herein.

[0069] In certain embodiments, the CDK2 inhibitors of the present disclosure (e.g., the compounds of Tables 1-3) are used in a method of treating a subject having, or at risk of developing, a disease or disorder associated with CDK2. The method can include administering a therapeutically effective amount of a compound described herein, or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition of a compound described herein, or the pharmaceutical dosage form of a compound described herein.

[0070] In certain embodiments, the CDK2 inhibitors of the present disclosure (e g., the compounds of Tables 1-3) are used in a method of treating a CDK2-mediated disorder or disease in a subject. The method can include administering a therapeutically effective amount of a compound described herein, or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition of a compound described herein, or the pharmaceutical dosage form of a compound described herein.

[0071] As used herein, the term “CDK2-mediated” disorders, diseases, and / or conditions means any disease or other deleterious condition in which CDK2 or a mutant thereof is known to play a role. Accordingly, another embodiment of the present disclosure relates to treating or lessening the severity of one or more diseases in which CDK2, or a mutant thereof, is known to play a role. Such CDK2-mediated disorders include but are not limited to proliferative disorders (e g. cancer).

[0072] In certain embodiments, the CDK2 inhibitors of the present disclosure (e.g., the compounds as described in any of the above embodiments) can be useful in applications that benefit from inhibition of CDK2 enzymes. In addition, CDK2 inhibitors described herein can be useful for the treatment of proliferative diseases generally.

[0073] The term “cyclin E” comprises two separate gene products, termed cyclin El and cyclin E2. Cyclin El is a well-established oncogene. The CCNE1 gene encoding cyclin El is amplified in many cancers, including uterine, ovarian, triple-negative breast, and esophageal cancers. In addition, cyclin El can be overexpressed due to oncogenic growth factor signaling leading to upregulation of CCNE1 transcription, or loss of function of tumor suppressor genes such as RBI, which increases CCNE1 transcription, or FBXW7, which normally mediates cyclin El degradation. Loss of FBXW7 function increases cyclin El stability and causes cyclin El accumulation (Sailo et al. Cancers (2019) 11 :246-276). Cyclin E2 is also overexpressed in many cancer types, including luminal breast cancers, via mechanisms similar to those described for cyclin El, including increased transcription and decreased degradation (Milioli et al. Endocrine Related Cancer (2020) 27: R93-R112).

[0074] Overexpression of either cyclin El or E2 drives hyperactivation of CDK2, which leads to dysregulated cell proliferation, genetic instability, and tumor growth and metastasis. After the initiation of DNA replication, CDK2 can also associate with cyclin Al or cyclin A2, and triggers responses that prepare the cell for mitotic cell division (De Boer et al., Oncogene (2008)27:4261-4268). A CDK2 inhibitor is expected to suppress CDK2 functions when it is associated with either the E-type or A-type cyclins.

[0075] CDK2 is known to be a factor in tumorigenesis and proliferation in many cancer types including, but not limited to, lung cancer, liver cancer, colon cancer, ovarian cancer, uterine (endometrial), and breast cancer (Opyrchal, Int J Oncol 2014; Shi, PLoS One 2015; Lim, Cancer Prev Res 2014). There is evidence showing that CDK2 is functionally linked with abnormal proliferation in multiple cancer types, and is therefore recognized as a potential therapeutic target for cancer therapy (Chohan, Curr Med Chem 2015). Persistent activation of CDK2 can drive the malignant transformation of breast epithelial cells. Suppression of CDK2 activity can effectively inhibit the proliferation of human breast cancer cells (Ali, Cancer Res 2009). Transgenic expression of constitutively-activated form of CDK2 in mouse mammary glands induces a highly invasive form of basal-like breast cancer (Corsino, Neoplasia 2008). Cyclin D1 / CDK2 complexes were detected in human breast cancer cell lines (Sweeney, Oncogene 1998), and the levels of these complexes correlated well with the degree of cyclin DI overexpression. The role of cyclin E and its associated kinase CDK2 in ovarian cancer has been investigated by screening primary, metastatic, recurrent and benign ovarian tumors. The CCNE1 gene encoding cyclin El was shown to be amplified in 21% and CDK2 in 6.4% of the cases analyzed. Additionally, cyclin El RNA was overexpressed in 29.5% and CDK2 in 6.5% of ovarian tumors tested. CDK2 expression has been found to be significantly elevated in glia-derived tumors especially in glioblastoma multiforme (GBM) and was functionally required for GBM cell proliferation and tumorigenesis (Wang, Transl Oncol 2016). CDK2 expression was significantly enriched in GBM tumors and functionally required for tumor proliferation both in vitro and in vivo. Additionally, high CDK2 expression was associated with poor prognosis in GBM patients. Radioresistance is a major predictor of poor clinical prognosis and tumor recurrence in GBM patients. CDK2 was found to be one of the most up-regulated kinase encoding genes in GBM after radiation treatment, and its expression level is correlated with the risk of disease recurrence (Wang, Transl Oncol 2016). Elevated levels of CDK2 expression have been observed in human cholangiocarcinoma, and transcriptional repression of CDK2 induced cell cycle arrest and restrained tumor growth (Zheng, Oncol Rep 2016). CDK2 overexpression in oral squamous cell carcinoma (SCC) may elevate pRB phosphorylation and permit more rapid entry of the cancer cells into S phase. In a clinicopathological survey of oral SCC, incidence of CDK2 expression was high in the poorlydifferentiated lesions, and was associated with tumor invasion, lymph node involvement and survival, an indication that change in CDK2 expression is associated with oral cancer progression and aggressiveness (Mihara, Jpn J Cancer Res 2001). Thus, increased expression of CDK2 is a factor in oral cancer progression and a negative predictive marker of the patients' prognosis (Mihara, Jpn J Cancer Res 2001). CDK2 plays a role in cell proliferation of non-small cell lung cancer (NSCLC) (Kawana, Am J Pathol 1998) or small cell lung cancer (SCLC). CDK2 has also been found to play a role in cell proliferation of prostate cancer (Flores, Endocrinology 2010).

[0076] Provided compounds (e.g., the compounds of Tables 1-3) are inhibitors of CDK2 and are therefore useful for treating one or more disorders associated with activity of CDK2 or mutants thereof. Thus, in certain embodiments, the present disclosure provides a method of treating a CDK2-mediated disorder in a subject comprising administering a therapeutically effective amount of a compound described herein, or a pharmaceutically acceptable salt thereof, or a pharmaceutically acceptable composition of either of the foregoing, to a subject in need thereof. In certain embodiments, the present disclosure provides a method of treating a CDK2- mediated disorder in a subject comprising administering a therapeutically effective amount of a compound described herein, or a pharmaceutically acceptable composition thereof, to a subject in need thereof.

[0077] The compounds disclosed herein (e.g., the compounds of Tables 1-3) inhibit CDK2 and therefore are useful for treating diseases for which CDK2 is dysregulated, such as cancer. The present disclosure provides a method of inhibiting CDK2 in a subject in need thereof, comprising administering to the subject an effective amount of a compound disclosed herein, a pharmaceutically acceptable salt thereof or a pharmaceutical composition disclosed herein.

[0078] In some embodiments, deregulated expression and activity of CDK2 binding partner cyclins A and E have been associated with a variety of cancer types including, without limitation, breast, lung, colorectal tumors, pancreatic and colon cancer. In some embodiments, the disease or disorder associated with CDK2 is N-myc amplified neuroblastoma ceils (see Molenaar, et al, Proc Natl Acad. Sci USA 106(31): 12968-12973); K-Ras mutant lung cancers (see Hu, S., et al., Mol Cancer Ther., 2015 14(11): 2576-85; K-Ras-driven pancreatic and colon cancers (see Lukasik, P et al., Int J Mol Sci. 2021 Mar; 22(6): 2935); and cancers with FBXW7 loss of function mutation and CCNE1 overexpression (see Takada, et al., Cancer Res, 201777(18). 4881-4893), all of which are incorporated herein in their entirety for all purposes.

[0079] Subjects “in need of inhibiting CDK2” are those having a disease for which a beneficial therapeutic effect can be achieved by inhibiting CDK2, e.g., a slowing in disease progression, alleviation of one or more symptoms associated with the disease or increasing the longevity of the subject in view of the disease. In some embodiments, the disease or disorder associated with CDK2 is an adenocarcinoma, carcinoma, or cystadenocarcinoma.

[0080] In some embodiments, compounds disclosed herein can have a beneficial therapeutic effect for treating a patient having an amplified expression level of CCNE1 and suffering from, or at risk of developing, cancer, e.g., a solid tumor, a blood cancer. Disclosed herein is a method of treating a patient comprising administering to the patient a therapeutically effective amount of a compound disclosed herein, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition disclosed herein.

[0081] An amplification of the CCNE1 gene and / or an expression level of CCNE1 that is higher than a control expression level of CCNE1 is indicative / predictive that a human subject having or at risk of developing a disease or disorder associated with CDK2 will respond to a CDK2 inhibitor. In some embodiments, the expression level of CCNE1 may be the level of CCNE1 mRNA. In other embodiments, the expression level of CCNE1 may be the level of CCNE1 protein.

[0082] Similar statements can be applied to the expression of cyclin E2. In normal cell cycles, E-type cyclins (cyclin El and cyclin E2) associate with CDK2 to promote Gl / S transition. Cyclin E / CDK2 complex mostly controls cell cycle progression and DNA replication through phosphorylation of specific substrates. Oncogenic activation of the Cyclin El or E2 / CDK2 complex impairs normal DNA replication, causing replication stress and DNA damage. As a consequence, Cyclin El or E2 / CDK2-induced replication stress leads to genomic instability and contributes to human carcinogenesis. For reasons that remain unclear, cyclin El overexpression appears to be more highly oncogenic than cyclin E2 overexpression. These and other indications suggest that cyclins El and E2 carry out both overlapping and distinct functions.

[0083] In some embodiments, the contemplated biomarker may be pl6 (also known as cyclin-dependent kinase inhibitor 2A, multiple tumor suppressor 1, and pl6INK4A), which acts as a negative regulator of the proliferation of normal cells by interacting with CDK4 or CDK6. In other embodiments, the contemplated biomarker may be phosphorylation of Rb at the serine corresponding to amino acid position 780. Rb is a regulator of the cell cycle and acts as a tumorsuppressor. Rb is activated upon phosphorylation by cyclin D-CDK4 / 6 at Ser780 and Ser795 and by cyclin E / CDK2 at Ser807 and Ser811.

[0084] The contemplated biomarker may also be selected from the group consisting of RBI, RBL1, RBL2, CDKN2A, CDKN1A, CDKN1B, FBXW7, CCNE1, CCNE2, CCNA1, CCNA2, CCND1, CCND2, CCND3, CDK2, CDK3, CDK4, CDK6, CDKN2A, CDNK1A, CDKN1B E2F1, E2F2, E2F3, C-MYC, MYCL, MYCN, EZH2, ER, HER2, HER3, HPV+, and EGFR.

[0085] The compounds disclosed herein inhibit CDK2 and therefore are useful for treating diseases for which CDK2 is dysregulated, such as cancer. The present disclosure provides a method of inhibiting CDK2 in a subject in need thereof, comprising administering to the subject an effective amount of a compound disclosed herein, a pharmaceutically acceptable salt thereof or a pharmaceutical composition disclosed herein.

[0086] In some embodiments, the disclosure provides a method of treating a disease or disorder associated with CDK2 in a patient, comprising administering to the patient a therapeutically effective amount of a compound of the present disclosure, or a pharmaceutically acceptable salt thereof.

[0087] In some embodiments, the disorder is a proliferative disorder. In some embodiments, the proliferative disorder is cancer. In some embodiments, the proliferative disorder is cancer, a tumor, malignancy, polyp, cyst, outgrowth, proliferative neoplastic, metastatic cancer, or a treatment-resistant cancer.

[0088] In some embodiments, the proliferative disorder is breast cancer (e.g. ER- positive / HR-positive, HER2-negative breast cancer, HER2-positive breast cancer, ER- positive / HR-positive, HER2 -positive breast cancer, triple negative breast cancer (TNBC), inflammatory breast cancer, ductal carcinoma in situ, inflammatory breast cancer, ductal carcinoma, Paget’s disease of the breast, invasive lobular carcinoma, lobular carcinoma in situ, metastatic breast cancer, phyllodes tumor, invasive tubular breast carcinoma, mucinous neoplasm, male breast cancer, angiosarcoma, metaplastic carcinoma, or medullary carcinoma), prostate cancer (e.g., acinar adenocarcinoma, prostatic ductal adenocarcinoma, or prostate sarcoma), lung cancer (e.g. lung nodules, non-small cell lung cancer, small cell lung cancer, mesothelioma, large cell lung cancer, salivary gland-type lung carcinoma, thymomas, or lung carcinoids), thyroid cancer (e.g. papillary thyroid cancer, follicular thyroid cancer, medullary thyroid cancer, Hurthlecell thyroid cancer, or anaplastic thyroid cancer), gastric cancer, ovarian cancer (e g. epithelial ovarian carcinomas, germ cell tumor, or stromal cell tumors), rectal cancer, endometrial carcinoma (e.g, endometrioid adenocarcinoma, uterine papillary serous carcinoma, uterine clear cell carcinoma,) and other uterine cancers (e.g., uterine carcinosarcoma or uterine sarcoma), anal cancer, bladder cancer (e.g. urothelial carcinoma, squamous cell bladder cancer, adenocarcinoma, sarcoma, or small cell bladder cancer), bone cancer (e.g. osteosarcoma, chondrosarcoma, Ewing sarcoma, fibrosarcoma, or malignant fibrous histiocytoma), brain cancer (e.g. glioma, meningioma, astrocytoma, glioblastoma multiforme, medulloblastoma, ependymoma, oligodendroglioma, craniopharyngioma, pituitary adenoma, Schwannoma, anaplastic astrocytoma, germ cell tumor, primitive neuroectodermal tumor, chordoma, haemangioblastoma, optic nerve glioma, subependymoma, or germinoma), cervical cancer (e.g. squamous cell carcinomas or cervical adenocarcinoma), colorectal cancer (e.g., adenocarcinoma, squamous cell carcinoma, and colon cancer), esophageal cancer (e.g. adenocarcinoma or squamous cell carcinoma, or small cell carcinoma), gastrointestinal stromal tumors, gestational trophoblastic disease (e.g. hydatidiform mole (HM), also called molar pregnancy, and gestational trophoblastic neoplasia), appendiceal cancer (e.g. epithelial appendiceal cancer or neuroendocrine appendiceal cancer), head and neck cancer (e.g. laryngeal and hypopharyngeal cancer, nasal cavity and paranasal sinus cancer, nasopharyngeal cancer, oral and oropharyngeal cancer, or salivary gland cancer), Hodgkin lymphoma (e.g. classical Hodgkin lymphoma or nodular lymphocyte- predominant Hodgkin lymphoma), non-Hodgkin lymphoma (e g. diffuse large B-cell lymphoma (DLBCL), follicular lymphoma, mantle cell lymphoma, small lymphocytic lymphoma, double hit / triple hit lymphoma, or primary mediastinal large B-cell lymphoma), Kaposi sarcoma (e.g. HIV-related Kaposi's sarcoma, classic Kaposi's sarcoma, transplant Kaposi's sarcoma, endemic, or African Kaposi's sarcoma), kidney cancer (e g. renal cell carcinoma, urothelial carcinoma also called transitional cell carcinoma, sarcoma, Wilms tumor, or lymphoma), leukemia (e.g. acute lymphocytic leukemia, chronic lymphocytic leukemia, chronic myeloid leukemia, acute myeloid leukemia, hairy cell leukemia, acute promyelocytic leukemia, T-cell prolymphocytic leukemia, lymphoid leukemia, large granular lymphocytic leukemia, b-cell prolymphocytic leukemia, myeloproliferative neoplasm, or mastocytosis), liver cancer (e.g. hepatocellular carcinoma or cholangiocarcinoma (cancer of the bile ducts), laryngeal cancer (e.g. laryngeal squamous cell carcinoma), malignant mesothelioma (e.g. malignant pleural mesothelioma or malignantperitoneal mesothelioma), multicentric Castleman disease (e.g. POEMS-associated MCD, HHV- 8-associated MCD, or idiopathic MCD), myeloproliferative neoplasms (e.g. chronic myelogenous leukemia, polycythemia vera, primary myelofibrosis (also called chronic idiopathic myelofibrosis), essential thrombocythemia, chronic neutrophilic leukemia, or chronic eosinophilic leukemia), pancreatic cancer (e.g. adenocarcinoma or neuroendocrine tumors), retinoblastoma (e.g. unilateral retinoblastoma, bilateral retinoblastoma, intraocular retinoblastoma, or extraocular retinoblastoma), rhabdomyosarcoma (e.g. embryonal rhabdomyosarcoma, alveolar rhabdomyosarcoma, botryoid rhabdomyosarcoma, or pleomorphic rhabdomyosarcoma), skin cancer (e.g. melanoma, basal cell carcinoma, squamous cell carcinoma, or Merkel cell cancer), soft tissue sarcoma (e.g. rhabdomyosarcoma, synovial sarcoma, liposarcoma, Ewing sarcoma, leiomyosarcoma, alveolar soft part sarcoma, fibrosarcoma, epithelioid sarcoma, undifferentiated pleomorphic sarcoma, dermatofibrosarcoma, Kaposi sarcoma, angiosarcoma, neurofibrosarcoma, gastrointestinal stromal tumor, clear cell sarcoma, desmoplastic small-round-cell tumor, low-grade fibromyxoid sarcoma, or solitary fibrous tumor), testicular cancer (e.g. seminoma and nonseminomatous germ cell tumors), vaginal cancer (e.g. squamous cell carcinoma, vaginal intraepithelial neoplasia, verrucous carcinoma, clear cell adenocarcinoma, papillary adenocarcinoma, mucinous adenocarcinoma, adenosquamous carcinoma, embryonal rhabdomyosarcoma, melanoma, small cell cancer of the vagina, or vaginal lymphoma), vulvar cancer (e.g. squamous cell carcinoma, verrucous carcinoma, adenocarcinoma, melanoma, or sarcoma), or neuroblastoma. In some embodiments, the proliferative disorder is intrahepatic cholangiocarcinoma, hepatocellular carcinoma, breast cancer, prostate cancer, lung squamous cell carcinoma, thyroid cancer, gastric cancer, or ovarian cancer. In some embodiments, the proliferative disorder is gastric cancer, hormone receptor-positive breast cancer, HER2-positive breast cancer, triple negative breast cancer, or rectal cancer. In some embodiments, the proliferative disorder is endometrial carcinoma, non-small cell lung cancer, lung squamous cell carcinoma, gastric cancer, breast cancer, or urothelial cancer.

[0089] In some embodiments, cancers treatable with compounds of the present disclosure include advanced / relapsed tumors, CCNE1 amplified platinum-resistant or platinum-refractory ovarian cancer or endometrial cancer (with prior platinum therapy) that has progressed following two or more lines of therapies; and gastric cancer (with prior platinum therapy) that has progressed following two or more lines of therapies; and ER+ HER2- breast cancer that has progressed despitetreatment with a CDK4 / 6 inhibitor. In some embodiments, cancers treatable with compounds of the present disclosure include platinum-resistant or platinum-refractory CCNE1 amplified ovarian cancer, CCNE1 amplified endometrial cancer that has failed two or more lines of therapies, CCNE1 amplified advanced / relapsed tumors that do not belong to the other groups; ER+ HER2- breast cancer that has progressed despite treatment with a CDK4 / 6 inhibitor; platinum-resistant or platinum- refractory CCNE1 amplified ovarian cancer; and ER+ HER2- breast cancer that has progressed despite treatment with a CDK4 / 6 inhibitor.Pharmaceutical Compositions

[0090] In some embodiments, the compounds of the present disclosure are administered as a pharmaceutical composition. When employed as pharmaceuticals, the compounds of the disclosure can be administered in the form of pharmaceutical compositions which is a combination of the compounds of the disclosure and a pharmaceutically acceptable carrier. These compositions can be prepared in a manner well known in the pharmaceutical art, and can be administered by a variety of routes. Such pharmaceutical compositions can be administered systemically. The term “systemic” as used herein includes parenteral, topical, transdermal, oral, by inhalation / pulmonary, rectal, nasal, buccal, and sublingual administration. The term “parenteral” as used herein includes subcutaneous, intradermal, intravenous, intramuscular, intracranial, and intraperitoneal administration. Preferably, the compounds are administered intramuscularly, subcutaneously, orally, or intranasally in therapeutically effective amounts to treat diseases. Any composition described herein can also be formulated for “non-systemic” or local oral, sublingual, cutaneous, percutaneous, subcutaneous, transdermal, intramuscular, intravenous, intravascular, or nasal, delivery. In some embodiments, oral delivery may include liquid drug products (e.g., elixirs, suspensions, solutions, syrups) that are packaged with dosage delivery devices (e.g., calibrated cups, droppers, syringes, spoons).

[0091] Provided herein, in certain embodiments, is a pharmaceutical composition comprising a therapeutically effective amount of any compound described herein. In certain instances, the pharmaceutical composition comprises a CDK2 inhibitor (e.g., any CDK2 inhibitor described herein).

[0092] In certain embodiments, pharmaceutical compositions are formulated in a conventional manner using one or more physiologically acceptable carriers including, e.g., excipients and auxiliaries which facilitate processing of the active compounds into preparationswhich are suitable for pharmaceutical use. In certain embodiments, proper formulation is dependent upon the route of administration chosen. A summary of pharmaceutical compositions described herein is found, for example, in Remington: The Science and Practice of Pharmacy, Nineteenth Ed (Easton, Pa.: Mack Publishing Company, 1995); Hoover, John E., Remington's Pharmaceutical Sciences, Mack Publishing Co., Easton, Pennsylvania 1975; Liberman, H.A. and Lachman, L., Eds., Pharmaceutical Dosage Forms, Mareel Decker, New York, N.Y., 1980; and Pharmaceutical Dosage Forms and Drug Delivery Systems, Seventh Ed. (Lippincott Williams & Wilkins 1999), all of which are incorporated herein in their entirety for all purposes.

[0093] A pharmaceutical composition, as used herein, refers to a mixture of a compound described herein, with other chemical components, such as carriers, stabilizers, diluents, dispersing agents, suspending agents, thickening agents, and / or excipients. In certain instances, the pharmaceutical composition facilitates administration of the compound to an individual or cell. In certain embodiments of practicing the methods of treatment or use provided herein, therapeutically effective amounts of compounds described herein are administered in a pharmaceutical composition to an individual having a disease, disorder, or condition to be treated. In specific embodiments, the individual is a human. As discussed herein, the compounds described herein are either utilized singly or in combination with one or more additional therapeutic agents.

[0094] In certain embodiments, the pharmaceutical formulations described herein are administered to an individual in any manner, including one or more of multiple administration routes, such as, by way of non-limiting example, oral, parenteral (e g., intravenous, subcutaneous, intramuscular), intranasal, buccal, topical, rectal, or transdermal administration routes.

[0095] In certain embodiments, a pharmaceutical composition described herein includes one or more compound described herein as an active ingredient in free-acid or free-base form, or in a pharmaceutically acceptable salt form. In some embodiments, the compounds described herein are utilized as an N-oxide or in a crystalline or amorphous form (i.e., a polymorph). In some situations, a compound described herein exists as tautomers. All tautomers are included within the scope of the compounds presented herein. In certain embodiments, a compound described herein exists in an unsolvated or solvated form, wherein solvated forms comprise any pharmaceutically acceptable solvent, e.g., water, ethanol, and the like. The solvated forms of the compounds presented herein are also considered to be described herein.

[0096] A “carrier” includes, in some embodiments, a pharmaceutically acceptable excipient and is selected on the basis of compatibility with compounds described herein, such as, e.g., a compound described in any of the above embodiments, and the release profile properties of the desired dosage form. Exemplary carrier materials include, e.g., binders, suspending agents, disintegration agents, filling agents, surfactants, solubilizers, stabilizers, lubricants, wetting agents, diluents, and the like. See, e.g., Remington: The Science and Practice of Pharmacy, Nineteenth Ed (Easton, Pa.: Mack Publishing Company, 1995); Hoover, John E., Remington's Pharmaceutical Sciences, Mack Publishing Co., Easton, Pennsylvania 1975; Liberman, H. A. and Lachman, L., Eds., Pharmaceutical Dosage Forms, Mareel Decker, New York, N.Y., 1980; and Pharmaceutical Dosage Forms and Drug Delivery Systems, Seventh Ed. (Lippincott Williams & Wilkins 1999), all of which are incorporated herein in their entirety for all purposes.

[0097] The pharmaceutical composition described herein optionally include an additional therapeutic compound described herein and one or more pharmaceutically acceptable additives such as a compatible carrier, binder, filling agent, suspending agent, flavoring agent, sweetening agent, disintegrating agent, dispersing agent, surfactant, lubricant, colorant, diluent, solubilizer, moistening agent, plasticizer, stabilizer, penetration enhancer, wetting agent, anti-foaming agent, antioxidant, preservative, or one or more combination thereof. In some aspects, using standard coating procedures, such as those described in Remington's Pharmaceutical Sciences, 20th Edition (2000), a film coating is provided around the formulation of the compound described in any of the above embodiments. In one embodiment, a compound described herein is in the form of a particle and some or all of the particles of the compound are coated. In certain embodiments, some or all of the particles of a compound described herein are microencapsulated. In some embodiments, the particles of the compound described herein are not microencapsulated and are uncoated.

[0098] Solid dosage forms for administration of the compounds according to the present disclosure can be manufactured by standard manufacturing techniques. Non-limiting examples of oral solid dosage forms for administration of the compounds of the disclosure are described below. Tablet and Capsule Compositions

[0099] Solid dosage forms for oral administration include capsules, tablets, pills, powders, and granules. In such solid dosage forms, the active compound is mixed with at least one inert, pharmaceutically acceptable excipient or carrier such as sodium citrate or dicalcium phosphate and / or (a) fillers or extenders such as starches, lactose, sucrose, glucose, mannitol, and silicic acid;(b) binders such as, for example, carboxymethylcellulose, alginates, gelatin, polyvinylpyrrolidinone, sucrose, and acacia; (c) humectants such as glycerol; (d) disintegrating agents such as agar-agar, calcium carbonate, potato or tapioca starch, alginic acid, certain silicates, and sodium carbonate, e) solution retarding agents such as paraffin; (f) absorption accelerators such as quaternary ammonium compounds; (g) wetting agents such as, for example, cetyl alcohol and glycerol monostearate; (h) absorbents such as kaolin and bentonite clay; and (i) lubricants such as talc, calcium stearate, magnesium stearate, solid polyethylene glycols, sodium lauryl sulfate, and mixtures thereof. In the case of capsules, tablets and pills, the dosage form may also comprise buffering agents.

[0100] Solid compositions of a similar type may also be employed as fillers in soft- and hard-filled gelatin capsules using such excipients as lactose or milk sugar as well as high molecular weight polyethylene glycols and the like. The solid dosage forms of tablets, dragees, capsules, pills, and granules can be prepared with coatings and shells such as enteric coatings and other coatings well known in the pharmaceutical formulating art. They may optionally contain opacifying agents and can also be of a composition that they release the active ingredient(s) only, or preferentially, in a certain part of the intestinal tract, optionally, in a delayed manner. Examples of embedding compositions that can be used include polymeric substances and waxes. Solid compositions of a similar type may also be employed as fillers in soft and hard-filled gelatin capsules using such excipients as lactose or milk sugar as well as high molecular weight polethylene glycols and the like.

[0101] The active compounds can also be in micro-encapsulated form with one or more excipients as noted above. The solid dosage forms of tablets, dragees, capsules, pills, and granules can be prepared with coatings and shells such as enteric coatings, release controlling coatings and other coatings well known in the pharmaceutical formulating art. In such solid dosage forms the active compound may be admixed with at least one inert diluent such as sucrose, lactose or starch. Such dosage forms may also comprise, as is normal practice, additional substances other than inert diluents, e.g., tableting lubricants and other tableting aids such a magnesium stearate and microcrystalline cellulose. In the case of capsules, tablets and pills, the dosage forms may also comprise buffering agents. They may optionally contain opacifying agents and can also be of a composition that they release the active ingredient(s) only, or preferentially, in a certain part of theintestinal tract, optionally, in a delayed manner. Examples of embedding compositions that can be used include polymeric substances and waxes.

[0102] The tablet or capsule compositions of the disclosure may be prepared according to techniques well-known in the art of pharmacy.

[0103] Tablets and capsule formulations may further contain one or more adjuvants, binders, diluents, disintegrants, excipients, fillers, or lubricants, each of which are known in the art. Examples of such include carbohydrates such as lactose or sucrose, dibasic calcium phosphate anhydrous, corn starch, mannitol, xylitol, cellulose or derivatives thereof, microcrystalline cellulose, gelatin, stearates, silicon dioxide, talc, sodium starch glycolate, acacia, flavoring agents, preservatives, buffering agents, disintegrants, and colorants. Orally administered compositions may contain one or more optional agents such as, e.g., sweetening agents such as fructose, aspartame or saccharin; flavoring agents such as peppermint, oil of wintergreen, or cherry; coloring agents; and preservative agents, to provide a pharmaceutically palatable preparation.Effervescent Compositions

[0104] The effervescent compositions of the disclosure may be prepared according to techniques well-known in the art of pharmacy.

[0105] Effervescent formulations contain and effervescent couple of a base component and an acid component, which components reach in the presence of water to generate a gas. In some embodiments, the base component may comprise, for example, an alkali metal or alkaline earth metal carbonate, or bicarbonate. The acid component may comprise, for example, an aliphatic carboxylic acid or a salt thereof, such as citric acid. The base and acid components may each independently constitute, for example, 25% to 55% (w / w) of the effervescent composition. The ratio of acid component to base component may be within the range of 1 : 2 to 2 : 1.

[0106] The effervescent compositions of the disclosure may be formulated using additional pharmaceutically acceptable carriers or excipients as appropriate. For example, one or more taste masking agents may be used. Dyes may also be used, as pediatric patients often prefer colorful pharmaceutical combinations. The compositions may take the form of, for example, tablets, granules or powders, granules or powders presented in a sachet.Chewable Tablets

[0107] The chewable tablets of the disclosure may be prepared according to techniques well-known in the art of pharmacy.

[0108] Chewable tablets are tablets that are intended to disintegrate in the mouth under the action of chewing or sucking and where, in consequence, the active ingredient has greater opportunity to come into contact with the bitter- taste receptors on the tongue.

[0109] One method of overcoming this issue is to absorb the active ingredient onto a suitable substrate. This approach is known in the art and described for example in U.S. Pat. No. 4,647,459, which is incorporated herein by reference in its entirety for all purposes.

[0110] Another approach involves forming the active ingredient into an aggregate along with a pre-swelled substantially anhydrous hydrocolloid. The hydrocolloid absorbs saliva and acquires a slippery texture which enables it to lubricate the particles of aggregate and mask the taste of the active ingredient. This approach is known in the art and described for example in European patent application 0190826, which is incorporated herein by reference in its entirety for all purposes. g

[0111] Another approach involves employing a water-insoluble hygroscopic excipient such as microcrystalline cellulose. This approach is known in the art and described for example in U.S. Pat. No. 5,275,823, which is incorporated herein by reference in its entirety for all purposes.

[0112] In addition to the above approaches, the chewable tablets of the present disclosure can also contain other standard tableting excipients such as a disintegrant and a taste-masking agent.Orodispersible Tablets

[0113] The orodispersible tablets of the disclosure may be prepared according to techniques well-known in the art of pharmacy.

[0114] In orodispersible tablets of the disclosure, the excipient mixtures are such as to provide it with a disintegration rate so that its disintegration in the buccal cavity occurs in an extremely short time and especially shorter than sixty seconds. In some embodiments, the excipient mixture is characterized by the fact that the active substance is in the form of coated or non-coated microcrystals of microgranules. In some embodiments, the orodispersible tablet comprises one or several disintegrating agents of the carboxymethylcellulose type or insoluble reticulated PVP type,one or several swelling agents which may comprise a carboxymethylcellulose, a starch, a modified starch, or a microcrystalline cellulose or optionally a direct compression sugar.Powders for Reconstitution

[0115] The powder for reconstitution pharmaceutical compositions of the disclosure may be prepared according to techniques well-known in the art of pharmacy.

[0116] In some embodiments, the powder for reconstitution compositions of the disclosure comprise an effective amount of at least one internal dehydrating agent. The internal dehydrating agent can enhance the stability of the powder. In some embodiments, the internal dehydrating agent is magnesium citrate or disodium carbonate. In some embodiments, the powder composition comprises a pharmaceutically acceptable diluents, such as sucrose, dextrose, mannitol, xylitol, or lactose.

[0117] Powder compositions of the disclosures may be placed in sachets or bottles for contemporaneous dissolution or for short term storage in liquid form (e g. 7 days).Gummy Candies

[0118] The gummy candies of the disclosure may be prepared according to techniques well-known in the art of pharmacy.

[0119] Traditional gummy candy is made from a gelatin base. Gelatin gives the candy its elasticity, the desired chewy consistency, and a longer shelflife. In some embodiments, the gummy candy pharmaceutical composition of the disclosure includes a binding agent, a sweetener, and an active ingredient.

[0120] In some embodiments, the binding agent is a pectin gel, gelatin, food starch, or any combination thereof.

[0121] In some embodiments, the gummy candy comprises sweeteners, a binding agent, natural and / or artificial flavors and colors and preservatives. In some embodiments, the gummy candy comprises glucose syrup, natural cane juice, gelatin, citric acid, lactic acid, natural colors, natural flavors, fractionated coconut oil, and carnauba wax.Liquid Dosage Forms

[0122] The pharmaceutical liquid dosage forms of the disclosure may be prepared according to techniques well-known in the art of pharmacy.

[0123] A solution refers to a liquid pharmaceutical formulation wherein the active ingredient is dissolved in the liquid. Pharmaceutical solutions of the disclosure include syrups and elixirs. A suspension refers to a liquid pharmaceutical formulation wherein the active ingredient is in a precipitate in the liquid.

[0124] In a liquid dosage form, it is desirable to have a particular pH and / or to be maintained within a specific pH range. In order to control the pH, a suitable buffer system can be used. In addition, the buffer system should have sufficient capacity to maintain the desired pH range. Examples of the buffer system useful in the present disclosure include but are not limited to, citrate buffers, phosphate buffers, or any other suitable buffer known in the art. Preferably the buffer system include sodium citrate, potassium citrate, sodium bicarbonate, potassium bicarbonate, sodium dihydrogen phosphate and potassium dihydrogen phosphate, etc. The concentration of the buffer system in the final suspension varies according to factors such as the strength of the buffer system and the pH / pH ranges required for the liquid dosage form. In one embodiment, the concentration is within the range of 0.005 to 0.5 w / v % in the final liquid dosage form.

[0125] The pharmaceutical composition comprising the liquid dosage form of the present disclosure can also include a suspending / stabilizing agent to prevent settling of the active material. Over time the settling could lead to caking of the active to the inside walls of the product pack, leading to difficulties with redispersion and accurate dispensing. Suitable stabilizing agents include but are not limited to, the polysaccharide stabilizers such as xanthan, guar and tragacanth gums as well as the cellulose derivatives HPMC (hydroxypropyl methylcellulose), methyl cellulose and Avicel RC-591 (microcrystalline cellulose / sodium carboxymethyl cellulose). In another embodiment, polyvinylpyrrolidone (PVP) can also be used as a stabilizing agent.

[0126] In addition to the aforementioned components, an oral liquid dosage form can also optionally contain other excipients commonly found in pharmaceutical compositions such as alternative solvents, taste-masking agents, antioxidants, fillers, acidifiers, enzyme inhibitors and other components as described in Handbook of Pharmaceutical Excipients, Rowe et al., Eds., 4thEdition, Pharmaceutical Press (2003), which is hereby incorporated by reference in its entirety for all purposes.

[0127] Addition of an alternative solvent may help increase solubility of an active ingredient in the liquid dosage form, and consequently the absorption and bioavailability insidethe body of a subject. Preferably the alternative solvents include methanol, ethanol or propylene glycol and the like.

[0128] In another aspect, the present disclosure provides a process for preparing the liquid dosage form. The process comprises steps of bringing the CDK2 inhibitor or its pharmaceutically acceptable salts thereof into mixture with the components including glycerol or syrup or the mixture thereof, a preservative, a buffer system and a suspending / stabilizing agent, etc., in a liquid medium. In general, the liquid dosage form is prepared by uniformly and intimately mixing these various components in the liquid medium. For example, the components such as glycerol or syrup or the mixture thereof, a preservative, a buffer system and a suspending / stabilizing agent, etc., can be dissolved in water to form the aqueous solution, then the active ingredient can be then dispersed in the aqueous solution to form a suspension.

[0129] In some embodiments, the liquid dosage form provided herein can be in a volume of between about 0.001 ml to about 100 ml. In some embodiments, the liquid dosage form provided herein can be in a volume of between about 0.1 ml to about 90 ml. In some embodiments, the liquid dosage form provided herein can be in a volume of between about 0.1 ml to about 80 ml. In some embodiments, the liquid dosage form provided herein can be in a volume of between about 0.1 ml to about 70 ml. In some embodiments, the liquid dosage form provided herein can be in a volume of between about 0.1 ml to about 60 ml. In some embodiments, the liquid dosage form provided herein can be in a volume of between about 0.1 ml to about 50 ml. In some embodiments, the liquid dosage form provided herein can be in a volume of between about 0.1 ml to about 40 ml. In some embodiments, the liquid dosage form provided herein can be in a volume of between about 0.1 ml to about 30 ml. In some embodiments, the liquid dosage form provided herein can be in a volume of between about 0.1 ml to about 20 ml. In some embodiments, the liquid dosage form provided herein can be in a volume of between about 0.1 ml to about 10 ml. In some embodiments, the liquid dosage form provided herein can be in a volume of between about 10 ml to about 30 ml. In some embodiments, the liquid dosage form provided herein can be in a volume of about 20 ml. In some embodiments, the CDK2 inhibitor can be in an amount ranging from about 0.001% to about 90% of the total volume. In some embodiments, the CDK2 inhibitor can be in an amount ranging from about 0.01% to about 80% of the total volume. In some embodiments, the CDK2 inhibitor can be in an amount ranging from about 0.1% to about 70% of the total volume. In some embodiments, the CDK2 inhibitor can be in an amount ranging from about 1% to about 60% ofthe total volume. In some embodiments, the CDK2 inhibitor can be in an amount ranging from about 1% to about 50% of the total volume. In some embodiments, the CDK2 inhibitor can be in an amount ranging from about 1% to about 40% of the total volume. In some embodiments, the CDK2 inhibitor can be in an amount ranging from about 1% to about 30% of the total volume. In some embodiments, the CDK2 inhibitor can be in an amount ranging from about 1% to about 20% of the total volume. In some embodiments, the CDK2 inhibitor can be in an amount ranging from about 1% to about 10% of the total volume. In some embodiments, the CDK2 inhibitor can be in an amount ranging from about 5% to about 70% of the total volume. In some embodiments, the CDK2 inhibitor can be in an amount ranging from about 5% to about 60% of the total volume. In some embodiments, the CDK2 inhibitor can be in an amount ranging from about 5% to about 50% of the total volume. In some embodiments, the CDK2 inhibitor can be in an amount ranging from about 5% to about 40% of the total volume. In some embodiments, the CDK2 inhibitor can be in an amount ranging from about 5% to about 30% of the total volume. In some embodiments, the CDK2 inhibitor can be in an amount ranging from about 5% to about 20% of the total volume. In some embodiments, the CDK2 inhibitor can be in an amount ranging from about 5% to about 10% of the total volume. In some embodiments, the CDK2 inhibitor can be in an amount ranging from about 10% to about 50% of the total volume. In some embodiments, the CDK2 inhibitor can be in an amount ranging from about 10% to about 40% of the total volume. In some embodiments, the CDK2 inhibitor can be in an amount ranging from about 10% to about 30% of the total volume. In some embodiments, the CDK2 inhibitor can be in an amount ranging from about 10% to about 20% of the total volume. In one embodiment, the resulted liquid dosage form can be in a liquid volume of 10 ml to 30 ml, preferably 20 ml, and the active ingredient can be in an amount ranging from about 0.001 mg / ml to about 16 mg / ml, or from about 0.025 mg / ml to about 8 mg / ml, or from about 0.1 mg / ml to about 4 mg / ml, or about 0.25 mg / ml, or about 0.5 mg / ml, or about 1 mg / ml, or about 2 mg / ml, or about 4 mg / ml, or about 5 mg / ml, or about 8 mg / ml, or about 10 mg / ml, or about 12 mg / ml, or about 14 mg / ml or about 16 mg / ml. In certain embodiments, tablets prepared for oral administration contain various excipients, including, by way of non-limiting example, binders, diluents, lubricants, disintegrants, fillers, stabilizers, surfactants, preservatives, coloring agents, flavoring agents and the like. In some embodiments, binders are used to impart cohesive qualities to a tablet, ensuring that the tablet remains intact after compression. Suitable binder materials include, by way of non-limiting example, starch (including corn starch and pregelatinized starch),gelatin, sugars (including sucrose, glucose, dextrose and lactose), polyethylene glycol, propylene glycol, waxes, and natural and synthetic gums, e.g., acacia sodium alginate, polyvinylpyrrolidone, cellulosic polymers (including hydroxypropyl cellulose, hydroxypropyl methylcellulose, methyl cellulose, ethyl cellulose, hydroxyethyl cellulose, and the like), Veegum, and combinations thereof. In certain embodiments, diluents are utilized to increase the bulk of the tablet so that a practical size tablet is provided. Suitable diluents include, by way of non-limiting example, dicalcium phosphate, calcium sulfate, lactose, cellulose, kaolin, mannitol, sodium chloride, dry starch, powdered sugar and combinations thereof. In certain embodiments, lubricants are used to facilitate tablet manufacture; examples of suitable lubricants include, by way of non-limiting example, vegetable oils such as peanut oil, cottonseed oil, sesame oil, olive oil, corn oil, and oil of theobroma, glycerin, magnesium stearate, calcium stearate, stearic acid and combinations thereof. In some embodiments, disintegrants are used to facilitate disintegration of the tablet, and include, by way of non-limiting example, starches, clays, celluloses, algins, gums, crosslinked polymers and combinations thereof. Fillers include, by way of non-limiting example, materials such as silicon dioxide, titanium dioxide, alumina, talc, kaolin, powdered cellulose and microcrystalline cellulose, as well as soluble materials such as mannitol, urea, sucrose, lactose, dextrose, sodium chloride and sorbitol. In certain embodiments, stabilizers are used to inhibit or retard drug decomposition reactions that include, by way of example, oxidative reactions. In certain embodiments, surfactants are anionic, cationic, amphoteric or nonionic surface active agents.Dosage Forms

[0130] Moreover, in certain embodiments, the pharmaceutical compositions described herein are formulated as a dosage form. As such, in some embodiments, provided herein is a dosage form comprising a compound described herein, suitable for administration to an individual. In certain embodiments, suitable dosage forms include, by way of non-limiting example, aqueous oral dispersions, liquids, gels, syrups, elixirs, slurries, suspensions, solid oral dosage forms, aerosols, controlled release formulations, fast melt formulations, effervescent formulations, lyophilized formulations, tablets, powders, pills, dragees, capsules, delayed release formulations, extended release formulations, pulsatile release formulations, multiparticulate formulations, and mixed immediate release and controlled release formulations.

[0131] In certain aspects, the composition or formulation containing one or more compounds described herein is orally administered for local delivery of a CDK2 inhibitor, or a compound described herein. Unit dosage forms of such compositions include a pill, tablet or capsules formulated. In certain embodiments, such pills, tablets or capsule contain the compositions described herein entrapped or embedded in microspheres. In some embodiments, microspheres include, by way of non-limiting example, chitosan microcores HPMC capsules and cellulose acetate butyrate (CAB) microspheres. In certain embodiments, oral dosage forms are prepared using conventional methods known to those in the field of pharmaceutical formulation. For example, in certain embodiments, tablets are manufactured using standard tablet processing procedures and equipment. An exemplary method for forming tablets is by direct compression of a powdered, crystalline or granular composition containing the active agent(s), alone or in combination with one or more carriers, additives, or the like. In alternative embodiments, tablets are prepared using wet-granulation or dry-granulation processes. In some embodiments, tablets are molded rather than compressed, starting with a moist or otherwise tractable material.

[0132] In other embodiments, the compositions described herein are administered orally for non-systemic delivery of the CDK2 inhibitor. In specific embodiments, compositions formulated for oral administration are, by way of non-limiting example, enterically coated or formulated oral dosage forms, such as, tablets and / or capsules.

[0133] In prophylactic applications, compounds or compositions containing compounds described herein may be administered to an individual susceptible to or otherwise at risk of a particular disease, disorder or condition. In certain embodiments of this use, the precise amounts of compound administered depend on the individual's state of health, weight, age, gender, and the like. Furthermore, in some instances, when a compound or composition described herein is administered to an individual, effective amounts for this use depend on the severity and course of the disease, disorder or condition, previous therapy, the individual's health status and response to the drugs, and the judgment of the treating physician.

[0134] In certain embodiments of the methods of the present disclosure, wherein following administration of a selected dose of a compound or composition described herein, an individual's condition does not improve, upon the doctor's discretion the administration of a compound or composition described herein is optionally administered chronically, that is, for an extended periodof time, including throughout the duration of the individual's life in order to ameliorate or otherwise control or limit the symptoms of the individual's disorder, disease or condition.

[0135] In certain embodiments of the methods of the present disclosure, an effective amount of a given agent varies depending upon one or more of a number of factors such as the particular compound, disease or condition and its severity, the identity (e.g., weight) of the subject or host in need of treatment, and is determined according to the particular circumstances surrounding the case, including, e.g., the specific agent being administered, the route of administration, the condition being treated, and the subject or host being treated. In some embodiments, doses administered include those up to the maximum tolerable dose. In some embodiments, doses administered include those up to the maximum tolerable dose by a newborn or an infant.

[0136] In various embodiments of the methods of the present disclosure, a desired dose is conveniently presented in a single dose or in divided doses administered simultaneously (or over a short period of time) or at appropriate intervals, for example as two, three, four or more subdoses per day. In various embodiments, a single dose of a CDK2 inhibitor is administered every 6 hours, every 12 hours, every 24 hours, every 48 hours, every 72 hours, every 96 hours, every 5 days, every 6 days, or once a week. In some embodiments the total single dose of a CDK2 inhibitor is in a range described below.

[0137] In various embodiments of methods of the present disclosure, in the case wherein the patient's status does improve, upon the doctor's discretion a CDK2 inhibitor is optionally given continuously; alternatively, the dose of drug being administered is temporarily reduced or temporarily suspended for a certain length of time (i.e., a “drug holiday”). The length of the drug holiday optionally varies between 2 days and 1 year, including by way of example only, 2 days, 3 days, 4 days, 5 days, 6 days, 7 days, 10 days, 12 days, 15 days, 20 days, 28 days, 35 days, 50 days, 70 days, 100 days, 120 days, 150 days, 180 days, 200 days, 250 days, 280 days, 300 days, 320 days, 350 days, or 365 days. The dose reduction during a drug holiday includes from 10%-100% of the original dose, including, by way of example only, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100% of the original dose. In some embodiments the total single dose of a CDK2 inhibitor is in a range described below.

[0138] Once improvement of the patient's conditions has occurred, a maintenance dose is administered if necessary. Subsequently, the dosage or the frequency of administration, or both, isreduced, as a function of the symptoms, to a level at which the improved disease, disorder or condition is retained. In some embodiments, patients require intermittent treatment on a long-term basis upon any recurrence of symptoms.

[0139] In certain instances, there are a large number of variables in regard to an individual treatment regime, and considerable excursions from these recommended values are considered within the scope described herein. Dosages described herein are optionally altered depending on a number of variables such as, by way of non-limiting example, the activity of the compound used, the disease or condition to be treated, the mode of administration, the requirements of the individual subject, the severity of the disease or condition being treated, and the judgment of the practitioner.

[0140] Toxicity and therapeutic efficacy of such therapeutic regimens are optionally determined by pharmaceutical procedures in cell cultures or experimental animals, including, but not limited to, the determination of the LDso (the dose lethal to 50% of the population) and the ED?o (the dose therapeutically effective in 50% of the population). The dose ratio between the toxic and therapeutic effects is the therapeutic index and it can be expressed as the ratio between LD50 and ED50. Compounds exhibiting high therapeutic indices are preferred. In certain embodiments, data obtained from cell culture assays and animal studies are used in formulating a range of dosage for use in human. In specific embodiments, the dosage of compounds described herein lies within a range of circulating concentrations that include the ED50 with minimal toxicity. The dosage optionally varies within this range depending upon the dosage form employed and the route of administration utilized.

[0141] The therapeutic dosage of the compounds of the disclosure can vary according to, for example, the particular use for which the treatment is made, the manner of administration of the compound, the health and condition of the patient, and the judgment of the prescribing physician. The proportion or concentration of the compounds of the disclosure in a pharmaceutical composition can vary depending upon a number of factors including dosage, chemical characteristics (e.g., hydrophobicity), and the route of administration. The dosage is likely to depend on such variables as the type and extent of progression of the disease or disorder, the overall health status of the particular patient, the relative biological efficacy of the compound selected, formulation of the excipient, and its route of administration. Effective doses can be extrapolated from dose-response curves derived from in vitro or animal model test systems.

[0142] In some embodiments, the compound of the disclosure is present in a composition or a unit dose of a composition described herein in an amount of from about 0.0001 mg to about 10,000 mg (e.g., about 0.001-1000 mg, about 0.01-100 mg, about 0.1-10 mg, about 1-5 mg or about 2-3 mg).

[0143] In certain aspects, about 0.05 mg to about 50 mg, about 0.25 mg to about 20 mg, about 0.25 mg to about 15 mg, about 0.25 mg to about 10 mg, or about 0.25 mg to about 5 mg (e.g., about 0.1 to about 5 mg, about 0.25 to about 2.5 mg, about 0.3 mg to about 2 mg, about 0.5 mg to about 1 mg, about 0.7 mg to about 1.5 mg, about 0.375 mg, about 0.75 mg, about 1 mg, about 1.25 mg, about 1.5 mg or about 2 mg) of the compound per day or per dose is administered to a patient.

[0144] In some embodiments, the compound is present in a unit dose in an amount of between about 5 mg and about 500 mg. In some embodiments, the amount of the compound administered daily or in a unit dose is between about 5 mg and about 300 mg. In other embodiments, the amount of the compound present in a unit dose or administered daily is between about 5 and about 250 mg, or between about 5 and about 200 mg, between about 5 mg and about 150 mg, between about 5 mg and about 100 mg, or between about 5 and about 50 mg.

[0145] In preparing a formulation, the active compound can be milled to provide the appropriate particle size prior to combining with the other ingredients. If the active compound is substantially insoluble, it can be milled to a particle size of less than 200 mesh. If the active compound is substantially water soluble, the particle size can be adjusted by milling to provide a substantially uniform distribution in the formulation, e.g. about 40 mesh. Some examples of suitable excipients include lactose, dextrose, sucrose, sorbitol, mannitol, starches, gum acacia, calcium phosphate, alginates, tragacanth, gelatin, calcium silicate, microcrystalline cellulose, polyvinylpyrrolidone, cellulose, water, syrup, and methyl cellulose. The formulations can additionally include, without limitation, lubricating agents such as talc, magnesium stearate, and mineral oil; wetting agents; emulsifying and suspending agents; preserving agents such as methyl- and propylhydroxy-benzoates; sweetening agents; and flavoring agents. The compositions of the disclosure can be formulated so as to provide quick, sustained or delayed release of the active ingredient after administration to the patient by employing procedures known in the art.

[0146] For preparing solid compositions such as tablets, the principal active ingredient is mixed with a pharmaceutical excipient to form a solid pre-formulation composition containing ahomogeneous mixture of a compound according to the disclosure. When referring to these preformulation compositions as homogeneous, the active ingredient is typically dispersed evenly throughout the composition so that the composition can be readily subdivided into equally effective unit dosage forms such as tablets, pills and capsules. This solid pre-formulation is then subdivided into unit dosage forms of the type described above containing from, for example, 0.000001 to about 2000 mg of the active ingredient of the present application.

[0147] The tablets or pills containing a compound according to the disclosure can be coated or otherwise compounded to provide a dosage form affording the advantage of prolonged action. For example, the tablet or pill can comprise an inner dosage and an outer dosage component, the latter being in the form of an envelope over the former. The two components can be separated by an enteric layer which serves to resist disintegration in the stomach and permit the inner component to pass intact into the duodenum or to be delayed in release. A variety of materials can be used for such enteric layers or coatings, such materials including a number of polymeric acids and mixtures of polymeric acids with such materials as shellac, cetyl alcohol, and cellulose acetate.

[0148] In some embodiments, the CDK2 inhibitor is administered at a dose of about or at least about 0.5 pg / kg, 1 pg / kg, 2 pg / kg, 3 pg / kg, 4 pg / kg, 5 pg / kg, 6 pg / kg, 7 pg / kg, 8 pg / kg, 9 pg / kg, 10 pg / kg, 15 pg / kg, 20 pg / kg, 25 pg / kg, 30 pg / kg, 35 pg / kg, 40 pg / kg, 45 pg / kg, 50 pg / kg, 55 pg / kg, 60 pg / kg, 65 pg / kg, 70 pg / kg, 75 pg / kg, 80 pg / kg, 85 pg / kg, 90 pg / kg, 100 pg / kg, 140 pg / kg, 150 pg / kg, 200 pg / kg, 240 pg / kg, 280 pg / kg, 300 pg / kg, 250 pg / kg, 280 pg / kg, 300 pg / kg, 400 pg / kg, 500 pg / kg, 560 pg / kg, 600 pg / kg, 700 pg / kg, 800 pg / kg, 900 pg / kg, 1,000 pg / kg, 1,100 pg / kg, 1,200 pg / kg, 1,300 pg / kg, 1,400 pg / kg, 1,500 pg / kg, 1,600 pg / kg, 1,700 pg / kg, 1,800 pg / kg, 1,900 pg / kg, or 2,000 pg / kg.

[0149] In various embodiments, the CDK2 inhibitor is administered at a dose not exceeding about 1 pg / kg, 2 pg / kg, 3 pg / kg, 4 pg / kg, 5 pg / kg, 6 pg / kg, 7 pg / kg, 8 pg / kg, 9 pg / kg, 10 pg / kg, 15 pg / kg, 20 pg / kg, 25 pg / kg, 30 pg / kg, 35 pg / kg, 40 pg / kg, 45 pg / kg, 50 pg / kg, 55 pg / kg, 60 pg / kg, 65 pg / kg, 70 pg / kg, 75 pg / kg, 80 pg / kg, 85 pg / kg, 90 pg / kg, 100 pg / kg, 140 pg / kg, 150 pg / kg, 200 pg / kg, 240 pg / kg, 280 pg / kg, 300 pg / kg, 250 pg / kg, 280 pg / kg, 300 pg / kg, 400 pg / kg, 500 pg / kg, 560 pg / kg, 600 pg / kg, 700 pg / kg, 800 pg / kg, 900 pg / kg, 1,000 pg / kg, 1,100 pg / kg, 1,200 pg / kg, 1,300 pg / kg, 1,400 pg / kg, 1,500 pg / kg, 1,600 pg / kg, 1,700 pg / kg, 1,800 pg / kg, 1,900 pg / kg, 2,000, or 2, 100 pg / kg.

[0150] In various embodiments, the CDK2 inhibitor is administered at a dose of about or of at least about 0.5 mg / day, 1 mg / day, 2 mg / day, 3 mg / day, 4 mg / day, 5 mg / day, 6 mg / day, 7 mg / day, 8 mg / day, 9 mg / day, 10 mg / day, 11 mg / day, 12 mg / day, 13 mg / day, 14 mg / day, 15 mg / day, 16 mg / day, 17 mg / day, 18 mg / day, 19 mg / day, 20 mg / day, 30 mg / day, 40 mg / day, 50 mg / day, 60 mg / day, 70 mg / day, 80 mg / day, 90 mg / day, 100 mg / day, 150 mg / day, 200 mg / day, 300 mg / day, 500 mg / day, 600 mg / day, 700 mg / day, 800 mg / day, 900 mg / day, or 1000 mg / day.

[0151] In various embodiments, the CDK2 inhibitor is administered at a dose of not more than about 1 mg / day, 2 mg / day, 3 mg / day, 4 mg / day, 5 mg / day, 6 mg / day, 7 mg / day, 8 mg / day, 9 mg / day, 10 mg / day, 11 mg / day, 12 mg / day, 13 mg / day, 14 mg / day, 15 mg / day, 16 mg / day, 17 mg / day, 18 mg / day, 19 mg / day, 20 mg / day, 30 mg / day, 40 mg / day, 50 mg / day, 60 mg / day, 70 mg / day, 80 mg / day, 90 mg / day, 100 mg / day, 150 mg / day, 200 mg / day, 300 mg / day, 500 mg / day, 600 mg / day, 700 mg / day, 800 mg / day, 900 mg / day, 1,000 mg / day, or 1,100 mg / day.

[0152] In some embodiments, the CDK2 inhibitor is administered at a dose of from about 140 gg / kg / day to about 1400 gg / kg / day. In various embodiments, the CDK2 inhibitor is administered at a dose of about or at least about 0.5 gg / kg / day, 1 gg / kg / day, 2 gg / kg / day, 3 gg / kg / day, 4 gg / kg / day, 5 gg / kg / day, 6 gg / kg / day, 7 gg / kg / day, 8 gg / kg / day, 9 gg / kg / day, 10 gg / kg / day, 15 gg / kg / day, 20 gg / kg / day, 25 gg / kg / day, 30 gg / kg / day, 35 gg / kg / day, 40 gg / kg / day, 45 gg / kg / day, 50 gg / kg / day, 100 gg / kg / day, 140 gg / kg / day, 150 gg / kg / day, 200 gg / kg / day, 240 gg / kg / day, 280 gg / kg / day, 300 gg / kg / day, 350 gg / kg / day, 380 gg / kg / day, 400 gg / kg / day, 500 gg / kg / day, 560 gg / kg / day, 600 gg / kg / day, 700 gg / kg / day, 800 gg / kg / day, 900 gg / kg / day, 1,000 gg / kg / day, 1,100 gg / kg / day, 1,200 gg / kg / day, or 1,300 gg / kg / day.

[0153] In various embodiments, the CDK2 inhibitor is administered at a dose not exceeding about 1 gg / kg / day, 2 gg / kg / day, 3 gg / kg / day, 4 gg / kg / day, 5 gg / kg / day, 6 gg / kg / day, 7 gg / kg / day, 8 gg / kg / day, 9 gg / kg / day 10 gg / kg / day, 15 gg / kg / day, 20 gg / kg / day, 25 gg / kg / day, 30 gg / kg / day, 35 gg / kg / day, 40 gg / kg / day, 45 gg / kg / day, 50 gg / kg / day, 100 gg / kg / day, 140 gg / kg / day, 150 gg / kg / day, 200 gg / kg / day, 240 gg / kg / day, 280 gg / kg / day, 300 gg / kg / day, 350 gg / kg / day, 380 gg / kg / day, 400 gg / kg / day, 500 gg / kg / day, 560 gg / kg / day, 600 gg / kg / day, 700 gg / kg / day, 800 gg / kg / day, 900 gg / kg / day, 1,000 gg / kg / day, 1,100 gg / kg / day, 1,200 gg / kg / day,1,300 gg / kg / day, or 1,400 gg / kg / day.

[0154] In various embodiments, the CDK2 inhibitor is administered at a dose of from about0.5 gg / kg / day to about 500 gg / kg / day, from about 0.5 gg / kg / day to about 250 gg / kg / day, fromabout 1 gg / kg / day to about 100 gg / kg / day, from about 10 gg / kg / day to about 50 gg / kg / day, from about 10 gg / kg / day to about 100 gg / kg / day, from about 0.5 gg / kg / day to about 2000 gg / kg / day, from about 280 gg / kg / day to about 1400 gg / kg / day, from about 420 gg / kg / day to about 1400 gg / kg / day, from about 250 to about 550 gg / kg / day, from about 560 gg / kg / day to about 1400 gg / kg / day, from 700 gg / kg / day to about 1400 gg / kg / day, from about 560 gg / kg / day to about 1200 gg / kg / day, from about 700 gg / kg / day to about 1200 gg / kg / day, from about 560 gg / kg / day to about 1000 gg / kg / day, from about 700 gg / kg / day to about 1000 gg / kg / day, from about 800 gg / kg / day to about 1000 gg / kg / day, from about 200 gg / kg / day to about 600 gg / kg / day, from about 300 gg / kg / day to about 600 gg / kg / day, from about 400 gg / kg / day to about 500 gg / kg / day, from about 400 gg / kg / day to about 600 gg / kg / day, from about 400 gg / kg / day to about 700 gg / kg / day, from about 400 gg / kg / day to about 800 gg / kg / day, from about 500 gg / kg / day to about 800 gg / kg / day, from about 500 gg / kg / day to about 900 gg / kg / day, from about 600 gg / kg / day to about 900 gg / kg / day, from about 700 gg / kg / day to about 900 gg / kg / day, from about 200 gg / kg / day to about 600 gg / kg / day, from about 800 gg / kg / day to about 900 gg / kg / day, from about 100 gg / kg / day to about 1500 gg / kg / day, from about 300 gg / kg / day to about 2,000 gg / kg / day, or from about 400 gg / kg / day to about 2000 gg / kg / day.

[0155] In some embodiments, the CDK2 inhibitor is administered at a dose of from about 30 gg / kg to about 1400 gg / kg per dose. In some embodiments, the CDK2 inhibitor is administered at a dose of from about 0.5 gg / kg to about 2000 gg / kg per dose, from about 0.5 gg / kg to about 1500 gg / kg per dose, from about 100 gg / kg to about 700 gg / kg per dose, from about 5 gg / kg to about 100 gg / kg per dose, from about 10 gg / kg to about 500 gg / kg per dose, from about 50 gg / kg to about 1400 gg / kg per dose, from about 300 gg / kg to about 2,000 gg / kg per dose, from about 60 gg / kg to about 1200 gg / kg per dose, from about 70 gg / kg to about 1000 gg / kg per dose, from about 70 gg / kg to about 700 gg / kg per dose, from 80 gg / kg to about 1000 gg / kg per dose, from 80 gg / kg to about 800 gg / kg per dose, from 100 gg / kg to about 800 gg / kg per dose, from 100 gg / kg to about 600 gg / kg per dose, from 150 gg / kg to about 700 gg / kg per dose, from 150 gg / kg to about 500 gg / kg per dose, from 200 gg / kg to about 400 gg / kg per dose, from 200 gg / kg to about 300 gg / kg per dose, or from 300 gg / kg to about 400 gg / kg per dose.

[0156] In some embodiments, the CDK2 inhibitor is administered at a dose of from about 0.5 mg / day to about 550 mg / day. In various embodiments, the CDK2 inhibitor is administered at a dose of from about 1 mg / day to about 500 mg / day, from about 1 mg / day to about 300 mg / day,from about 1 mg / day to about 200 mg / day, from about 2 mg / day to about 300 mg / day, from about 2 mg / day to about 200 mg / day, from about 4 mg / day to about 300 mg / day, from about 4 mg / day to about 200 mg / day, from about 4 mg / day to about 150 mg / day, from about 5 mg / day to about 150 mg / day, from about 5 mg / day to about 100 mg / day, from about 5 mg / day to about 80 mg / day, from about 5 mg / day to about 50 mg / day, from about 5 mg / day to about 40 mg / day, from about 5 mg / day to about 30 mg / day, from about 5 mg / day to about 20 mg / day, from about 5 mg / day to about 15 mg / day, from about 10 mg / day to about 100 mg / day, from about 10 mg / day to about 80 mg / day, from about 10 mg / day to about 50 mg / day, from about 10 mg / day to about 40 mg / day, from about 10 mg / day to about 20 mg / day, from about 20 mg / day to about 100 mg / day, from about 20 mg / day to about 80 mg / day, from about 20 mg / day to about 50 mg / day, or from about 20 mg / day to about 40 mg / day, or from about 20 mg / day to about 30 mg / day.

[0157] In various embodiments, the dose of the CDK2 inhibitor is a first dose level. In various embodiments, the dose of the CDK2 inhibitor is a second dose level. In some embodiments, the second dose level is greater than the first dose level. In some embodiments, the second dose level is about or at least about 1.5, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 30, 40, 50, 60, 70, 80, 90 or 100 times or fold greater than the first dose level. In some embodiments, the second dose level is not in excess of about 1.5, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 30, 40, 50, 60, 70, 80, 90, 100, or 150 times or fold greater than the first dose level.

[0158] In various embodiments, the CDK2 inhibitor is administered once daily (QD) at one of the above doses or within one of the above dose ranges. In various embodiments, the CDK2 inhibitor is administered twice daily (BID) at one of the above doses or within one of the above dose ranges. In various embodiments, a CDK2 inhibitor dose is administered daily, every other day, twice a week, or once a week.

[0159] In various embodiments, the CDK2 inhibitor is administered regularly for a period of about or of at least about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 30, 40, 48, 50, 75, 100, 150, 200, 250, 300, 350, 400, 450, 500, 600, 700, or 800 weeks. In various embodiments, the CDK2 inhibitor is administered for not more than about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 30, 40, 48, 50, 75, 100, 150, 200, 250, 300, 350, 400, 450, 500, 600, 700, 800, or 1000 weeks. In various embodiments, the CDK2 inhibitor is administered regularly for a period of about or of at least about 0.5, 1, 1.5, 2, 3, 4, 5, 6, 7, 8, 9, or 10 years. In various embodiments, the CDK2 inhibitor is administered regularly for a period not in excess of about 0.5, 1, 1.5, 2, 3, 4, 5, 6, 7, 8, 9, 10, or 15 years.Kits

[0160] In another aspect, provided herein are kits containing a device for administration of a pre-filled a pharmaceutical composition comprising a CDK2 inhibitor, as described herein. In certain embodiments, kits contain a device for oral administration and a pharmaceutical composition as described herein. In certain embodiments the kits include prefilled sachet or bottle for oral administration.

[0161] In certain embodiments, the present disclosure provides one or more containers containing a pharmaceutical composition comprising a therapeutically effective amount of the compounds of the disclosure. Such kits can further include, if desired, one or more of various conventional pharmaceutical kit components, such as, for example, containers with one or more pharmaceutically acceptable carriers, additional containers, etc., as will be readily apparent to those skilled in the art. Instructions, either as inserts or as labels, indicating quantities of the components to be administered, guidelines for administration, and / or guidelines for mixing the components, can also be included in the kit.

[0162] Delivery devices are important not only for delivering the compounds of the disclosure, but also for providing an appropriate environment for storage. This would include protection from microbial contamination and chemical degradation. The device and formulation should be compatible so as to avoid potential leaching or adsorption. The delivery device (or its packaging) can be optionally provided with a label and / or with instructions for use indicating how the composition should be administered, e.g., orally or subcutaneously.Combinations

[0163] In certain embodiments, a pharmaceutical composition comprising a CDK2 inhibitor, as described herein, can further include one or more additional therapeutic agents. In certain embodiments, a CDK2 inhibitor or a pharmaceutical composition thereof, as described herein, can be administered in combination with one or more additional therapeutic agents. The CDK2 inhibitor or a pharmaceutical composition thereof, and the one or more additional therapeutic agents, as described herein, may be administered simultaneously or sequentially in any order and / or at any interval appropriate to achieve a desirable therapeutic outcome.

[0164] In some embodiments, the one or more additional therapeutic agents is selected from antibodies, antibody-drug conjugates, kinase inhibitors, immunomodulators, histone deacetylase inhibitors, and anti-neoplastic agents. In some embodiments, the one or moreadditional therapeutic agent is selected from the following agents, or a pharmaceutically acceptable salt thereof: BCR-ABL inhibitors (e.g. imatinib, inilotinib, nilotinib, dasatinib, bosutinib, ponatinib, bafetinib, danusertib, saracatinib, PF03814735); ALK inhibitors (see Dardaei et al, 2018, Nat Med.; 24(4):512-517) (e.g. crizotinib, NVP-TAE684, ceritinib, alectinib, brigatinib, entrecinib, lorlatinib); BRAF inhibitors (see Prahallad et al, 2015, Cell Rep. 12, 1978-1985) (e g. vemurafenib, dabrafenib); FGFR inhibitors (e.g. infigratinib, dovitinib, erdafitinib, BLU-554, AZD4547); FLT3 inhibitors (e.g. sunitinib, midostaurin, tanutinib, sorafenib, lestaurtinib, quizartinib, and crenolanib); MEK Inhibitors (see Fedele et al, 2018, BioRxiv 307876; Torres- Ayuso et al, 2018, Cancer Discov.8, 1210-1212; and Wong et al, 2016, Oncotarget.2016 Oct 4; 7(40): 65676-65695) (e.g. trametinib, cobimetinib, binimetinib, selumetinib); ERK inhibitors (e.g. ulixertinib, MK-8353, LY-3214996); VEGF receptor inhibitors (e.g. bevacizumab, axitinib, aflibercept, brivanib, motesanib, pasireotide, sorafenib); Tyrosine kinase inhibitors (e.g. erlotinib, linifanib, sunitinib, pazopanib); Epidermal growth factor receptor (EGFR) inhibitors: gefitinib, osimertinib, cetuximab, panitumumab); HER2 receptor inhibitors (e.g. trastuzumab, neratinib, lapatinib, lapatinib); MET inhibitors (e.g. crizotinib, cabozantinib); CD20 antibodies (e.g. rituximab, tositumomab, ofatumumab); DNA Synthesis inhibitors (e.g. capecitabine, gemcitabine, nelarabine, hydroxycarbamide); Antineoplastic agents (e.g. oxaliplatin, cisplatin); HER dimerization inhibitors (e.g. pertuzumab); Human Granulocyte colony-stimulating factor (G-CSF) modulators (e.g. fdgrastim); Immunomodulators (e g. afutuzumab, lenalidomide, thalidomide, pomalidomide); CD40 inhibitors (e.g. dacetuzumab); Pro-apoptotic receptor agonists (PARAs) (e g. dulanermin); Heat Shock Protein (HSP) inhibitors (e.g. tanespimycin (17-allylamino-17- desmethoxygeldanamycin); Hedgehog antagonists (e.g. vismodegib); Proteasome inhibitors (e.g. bortezomib); PI3K inhibitors (e.g. pictilisib, dactolisib, buparlisib, taselisib, idelalisib, duvelisib, umbralisib); Phospholipase A2 inhibitors (e.g. anagrelide); BCL-2 inhibitors (e.g. venetoclax); Aromatase inhibitors (e.g. exemestane, letrozole, anastrozole, faslodex, tamoxifen); Topoisomerase I inhibitors (e.g. irinotecan, topotecan); Topoisomerase II inhibitors (e.g. etoposide, teniposide); mTOR inhibitors (e.g. temsirolimus, ridaforolimus, everolimus, sirolimus); Osteoclastic bone resorption inhibitors (e.g. zoledronic acid); CD33 Antibody Drug Conjugates (e.g. gemtuzumab ozogamicin); CD22 Antibody Drug Conjugates (e.g. inotuzumab ozogamicin); CD20 Antibody Drug Conjugates (e.g. ibritumomab tiuxetan); Somatostain analogs (e.g. octreotide); Interleukin- 11 (IL-11) (e.g. oprelvekin); Synthetic erythropoietin (e.g. darbepoetinalfa); Receptor Activator for Nuclear Factor K B (RANK) inhibitors (e.g. denosumab); Thrombopoietin mimetic peptides (e.g. romiplostim); Cell growth stimulators (e.g. palifermin); Anti-Insulin-like Growth Factor-1 receptor (IGF-1R) antibodies (e.g. figitumumab); Anti-CSl antibodies (e.g. elotuzumab); CD52 antibodies (e.g. alemtuzumab); CTLA-4 inhibitors (e.g. tremelimumab, ipilimumab); PD1 inhibitors (e.g. nivolumab, pembrolizumab); an immunoadhesin fusion protein (e.g. pidilizumab, AMP-224); PDL1 inhibitors (e.g. MSB0010718C, YW243.55.S70, atezolizumab, MEDI-4736, Avelumab, BMS-936559); LAG-3 inhibitors (e.g. relatlimab); GITR agonists; GITR fusion proteins and anti-GITR antibodies; Histone deacetylase inhibitors (HDI) (e.g. voninostat); Anti-CTLA4 antibodies (e.g. tremelimumab, ipilimumab); Alkylating agents (e.g. temozolomide, dactinomycin, melphalan, altretamine carmustine, bendamustine, busulfan, carboplatin, lomustine, cisplatin, chlorambucil, cyclophosphamide, dacarbazine, altretamine, ifosfamide, procarbazine, mechlorethamine, mustine and mechloroethamine, streptozocin, thiotepa); Biologic response modifiers (e.g. bacillus calmette- guerin, denileukin diftitox); Anti-tumor antibiotics (e.g. doxorubicin, bleomycin, daunomycin, daunorubicin, liposomal mitoxantrone epirubicin, idarubicin, mitomycin C); Anti -microtubule agents (e.g. estramustine); Cathepsin K inhibitors (e.g. odanacatib); Epothilone analogs (e g. ixabepilone); TpoR agonists (e.g. eltrombopag); Anti-mitotic agents (e.g. docetaxel); Adrenal steroid inhibitors (e.g. aminoglutethimide); Anti-androgens (e.g. nilutamide); Androgen Receptor inhibitors (e.g. enzalutamide, abiraterone acetate, orteronel, galeterone, and seviteronel, bicalutamide, flutamide); Androgens (e g. fluoxymesterone); CDK inhibitors (e.g. alvocidib, palbociclib, ribociclib, trilaciclib, abemaciclib); Gonadotropin-releasing hormone (GnRH) receptor agonists (e.g. leuprolide or leuprolide acetate); Taxane anti-neoplastic agents (e.g. cabazitaxel, larotaxel); 5-HTla receptor agonists (e.g. xaliproden); HPV vaccines (e.g. Cervarix® sold by GlaxoSmithKline, Gardasil® sold by Merck); Iron Chelating agents (e.g. deferasirox); Anti-metabolites (e.g. cladribine, 5-fluorouracil, 6-thioguanine, pemetrexed, cytarabine, cytarabine liposomal, decitabine, hydroxyurea, fludarabine, floxuridine, cladribine, methotrexate, pentostatin); Bisphosphonates (e.g. pamidronate); Demethylating agents (e.g. 5-azacitidine, decitabine); Anti-tumor Plant Alkaloids (e.g. paclitaxel protein-bound, vinblastine, vincristine, vinorelbine, paclitaxel); Retinoids (e.g. alitretinoin, tretinoin, isotretinoin, bexarotene); Glucocorticosteroids (e.g. hydrocortisone, dexamethasone, prednisolone, prednisone, methylprednisolone); Cytokines (e.g. interleukin-2, interleukin- 11 (oprevelkin), alpha interferonalfa (IFN-alpha)); estrogen receptor downregulators (e.g. fulvestrant); Anti-estrogens (e g. tamoxifen, toremifene); Selective estrogen receptor modulators (SERMs) (e.g. raloxifene); Luteinizing hormone releasing hormone (LHRH) agonists (e.g. goserelin); Progesterones (e.g. megestrol); cytotoxic agents (e.g. arsenic trioxide, asparaginase (also known as L-asparaginase, Erwinia L- asparaginase); Anti-nausea drugs (e.g. NK-1 receptor antagonists (e.g. casopitant)); Cytoprotective agents (e.g. amifostine, leucovorin); and Immune checkpoint inhibitors.

[0165] The term "immune checkpoints" refers to a group of molecules on the cell surface of CD4 and CD8 T cells. Immune checkpoint molecules include, but are not limited to, Programmed Death 1 (PD- 1), Cytotoxic T-Lymphocyte Antigen 4 (CTLA-4), B7H1, B7H4, 0X40, CD 137 (4-1BB), CD40, and LAG3. Immunotherapeutic agents which can act as immune checkpoint inhibitors useful in the methods of the present disclosure, include, but are not limited to, inhibitors of PD-L1, PD-L2, CTLA4, TIM3, LAG3, VISTA, BTLA, TIGIT, LAIR1, CD 160, 2B4 and / or TGF beta receptor.

[0166] As used herein, anti-neoplastic agents are drugs or active agents that are used to treat cancer (e.g., anticancer drugs, chemotherapy, cytotoxic drugs, and the like). The anti- neoplastic agent can include, without limitation, an antibody, an antibody-drug conjugate (e.g., sacituzumab govitecan, mirvetuximab soravtansine, trastuzumab deruxtecan, enfortumab vedotin, or tisotuniab vedotin), a DNA synthesis inhibitor (e.g., capecitabine, gemcitabine, nelarabine, or hydroxycarbamide), a platinum-based agent (e.g., carboplatin, oxaliplatin or cisplatin), topoisomerase I inhibitor (e.g., irinotecan or topotecan), topoisomerase II inhibitor (e.g., etoposide or teniposide), an DNA-intercalating agent (e.g., doxorubicin or liposomal doxorubicin), an alkylating agent (e.g., temozolomide, dactinomycin, melphalan, altretamine, carmustine, bendamustine, busulfan, lomustine, chlorambucil, cyclophosphamide, dacarbazine, altretamine, ifosfamide, or procarbazine), an anti-microtubule agent (e.g., estramustine), a taxane-related antimitotic agent (e.g., docetaxel, paclitaxel, cabazitaxel, abraxane, or larotaxel), an anti-metabolite (e.g., cladribine, 5-fluorouracil, 6-thioguanine, pemetrexed, cytarabine, hydroxyurea, fludarabine, floxuridine, pentostatin, or methotrexate), an anti-tumor plant alkaloid (e.g., vinblastine, vincristine, or vinorelbine), an anti-estrogen agent (e.g., tamoxifen, toremifene, raloxifene, lasofoxifene, afimoxifene, arzoxifene, bazedoxifene, fispemifene, ormeloxifene, ospemifene, tesmilifene, trilostane, CHF 4227, fulvestrant, elacestrant, giredestrant, amcenestrant, camizestrant, ataraestane, formestane, exemestane, letrozole, anastrozole, fadrozole;gonadotropin-releasing hormone, leuprolide, leuprolide acetate, enzalutamide, abiraterone acetate, or bicalutamide), a protein kinase inhibitor (e.g., a CDK4 / 6 inhibitor (e.g, palbociclib, ribociclib, trilaciclib, or abemaciclib), alvocidib, crizotinib, NVP-TAE684, ceritinib, alectinib, brigatinib, entrectinib, lorlatinib, vemurafenib, dabrafenib, infigratinib, dovitinib, erdafitinib, BLU-554, AZD4547, trametinib, cobimetinib, binimetinib, selumetinib, ulixertinib, MK-8353, LY-3214966, bevacizumab, axitinib, aflibercept, brivanib, motesanib, pasireotide, sorafenib, erlotinib, linifanib, sunitinib, pazopanib, gefitinib, osimertinib, cetuximab, panitumumab, trastuzumab, neratinib, lapatinib, or cabozantinib), a phosphatidylinositol 3-kinase inhibitor (e.g., apelisib, duvelisib, copanlisib, or idelalisib), an immunomodulator, a histone deacetylase inhibitor (e.g., voninostat), a KRAS inhibitor (e.g., AMG510, MRTX849, JNJ-74699157 / ARS- 3248, Bl 1701963, Bl 1823911, BAY-293, GDC-6036, MRTX1133, a RAS(ON) inhibitor), or an immunotherapeutic agent (e.g, PD1, PD-L1, PD-L2, CTLA-4, LAG3, TIM3, VISTA, BTLA, TIGIT, LAIR1, CD160, 2B4, 4- IBB, GITR, 0X40, or TGF beta receptor), an epigenetic modulator (e.g, EZH2a, ARID1A, ARID2, PRMT1-9, BRIM, IDH1 / 2, or BCL6), and combinations thereof. In some embodiments, the immunotherapeutic agent is a CTLA-4 inhibitor (e.g, tremelimumab or ipilimumab), a PD1 inhibitor (e.g, nivolumab or pembrolizumab), a PD-L1 inhibitor (e.g, atezolizumab, avelumab, or durvalumab), a LAG3 inhibitor (e.g, comprises BMS-986016). In some embodiments, the immunotherapeutic agent comprises a CAR T-cell therapy.

[0167] In some embodiments, the one or more additional therapeutic agent is selected from the following agents: anti-CDK2 agents; cytotoxic agents; Estrogen Receptor-targeted or other endocrine therapies, immune-checkpoint inhibitors, other CDK inhibitors, Receptor Tyrosine Kinase inhibitors, BRAE inhibitors, MEK inhibitors, PI3K inhibitors, SHP2 inhibitors, and SRC inhibitors. (See M. Katoh, Nat. Rev. Clin. Oncol.2019, 16: 105-122; Y.K. Chae, et al. Oncotarget 2017, 8: 16052-16074; L. Formisano et al, Nat. Comm. 2019, 10: 1373-1386; and references cited therein, all of which are incorporated herein in their entirety for all purposes.)

[0168] In some embodiments, the additional anti -neoplastic agent may be selected from purine antagonists (such as 6-mercaptopurine riboside, mercaptopurine, 6-thioguanine, cladribine, clofarabine (Clolar), fludarabine, nelarabine, and raltitrexed), pyrimidine antagonists (such as 5- fluorouracil (5-FU), Alimta (pemetrexed di sodium, LY231514, MTA), capecitabine (Xeloda™), cvtosine arabinoside, gemcitabine (Gemzar™), Tegafur tUFT Orzel or Uforai and including TS-1 combination of tegafur, gimestat and otostat), doxifluridine, carmofur, cytarabine (includingocfosfate, phosphate stearate, sustained release and liposomal forms), enocitabine, 5-azac-itidine (Vidaza), decitabine, and ethynylcytidine), dihydrofolate reductase inhibitors (such as methotrexate and NeuTrexin (trimetresate glucuronaie)), and other anti metabolites such as AG- 014699, ABT-472, INO-lOOl, KU-0687, GPI 18180, eflornithine, hydroxyurea, leucovorin, nolatrexed (Thyrnitaq), triapine, trimetrexate, and N-(5-[N-(3,4-dihydro-2-methy1-4- oxoquinazolin-6-ylmethyl )-N-methylaminol”2-thenoyf)-L-glutamic acid, and combinations thereof

[0169] Further examples of anti-neoplastic agents include, but are not limited to, Abraxane, Advexin (ING 201), Batabulin, bexarotene (Targretin™), EPO 906, Vinflunine, actinomycin D, bleomycin, mitomycin C, neocarzi nostatin (Zinostatin), vinblastine, vincristine, vindesine, vinorelbine (Navelbine), docetaxel (Taxotere), Ortataxel, paclitaxel (including Taxoprexin a DHA / paclitaxel conjugate), Nedaplatin, oxaliplatin (Eloxatin), Satraplatin, Camptosar, capecitabine (Xeloda), oxaliplatin (Eloxatin), Taxotere alitretinoin, Canfosfamide (Telcyta™), DMXAA (Antisoma), ibandronic acid, L-asparaginase, pegaspargase (Oncaspar™), Efaproxiral (Efaproxyn™ - radiation therapy), Tesmilifene (DPPE-- enhances efficacy of cytotoxics), Theratope™ (Bioniira), Tretinoin (Vesanoid™), tirapazamine (Trizaone™), motexafm gadolinium (Xcytrin™) Cotara™ (mAb), and NB1-300L polyglutamate- paclitaxel (Xyotax™), TNFerade (GeneVec, a compound which express TNFalpha in response to radiotherapy), RB94, Genasense (Oblimersen), Combretastatin A4P (CA4P), Oxi -4503, AVE- 8062, TZT-1027, Atorvastatin (Lipitor), Provastatin (Pravachol), Lovastatin (Mevacor), Simvastatin (Zocor), Fluvastatin (Lescol), Ceiivastatin (Baycol), Rosuvastatin (Crestor), Lovostatin, Niacin (Advicor), Caduet, torcetrapib, ZD-6126, and combinations thereof.

[0170] A compound described herein (e.g., a compound described in any of the above embodiments) may also be used in combination with known therapeutic processes, for example, the administration of hormones or radiation. In certain embodiments, a provided compound is used as a radiosensitizer, especially for the treatment of tumors which exhibit poor sensitivity to radiotherapy.

[0171] A compound described herein (e.g., a compound described in any of the above embodiments) can be administered alone or in combination with one or more other therapeutic compounds, possible combination therapy taking the form of fixed combinations or the administration of a compound described herein and one or more other therapeutic compoundsbeing staggered or given independently of one another, or the combined administration of fixed combinations and one or more other therapeutic compounds. A compound described herein can besides or in addition be administered especially for tumor therapy in combination with chemotherapy, radiotherapy, immunotherapy, phototherapy, surgical intervention, or a combination of these. Long-term therapy is equally possible as is adjuvant therapy in the context of other treatment strategies, as described above. Other possible treatments are therapy to maintain the patient's status after tumor regression, or even chemopreventive therapy, for example in patients at risk.

[0172] In one embodiment, the compound described herein (e.g., the compounds of Tables 1-3) may be used in combination with a CDK4 / CDK6 inhibitor. The CDK4 / CDK6 may be selected from palbociclib, riboci clib, trilaciclib, or abemaciclib.

[0173] In one embodiment, the compound described herein (e.g., the compounds of Tables 1-3) may be used in combination with a chemotherapeutic agent. The chemotherapeutic agent may be selected from a DNA synthesis inhibitor (e.g., capecitabine, gemcitabine, nelarabine, or hydroxycarbamide), a platinum-based agent (e.g., carboplatin, oxaliplatin or cisplatin), topoisomerase I inhibitor (e g., irinotecan or topotecan), topoisomerase II inhibitor (e.g., etoposide or teniposide), an DNA-intercalating agent (e.g., doxorubicin or liposomal doxorubicin), an alkylating agent (e.g., temozolomide, dactinomycin, melphalan, altretamine, carmustine, bendamustine, busulfan, lomustine, chlorambucil, cyclophosphamide, dacarbazine, altretamine, ifosfamide, or procarbazine), an anti -microtubule agent (e.g., estramustine), a taxane-related antimitotic agent (e.g., docetaxel, paclitaxel, cabazitaxel, abraxane, or larotaxel), an anti-metabolite (e.g., cladribine, 5-fluorouracil, 6-thioguanine, pemetrexed, cytarabine, hydroxyurea, fludarabine, floxuridine, pentostatin, or methotrexate), or an anti-tumor plant alkaloid (e.g., vinblastine, vincristine, or vinorelbine).EXAMPLES

[0174] The following examples are provided by way of illustration. The examples should not be construed as limiting, as the examples merely provide specific understanding and practice of the embodiments and their various aspects.Example 1 - Synthesis of Compounds Formula (I)-(IT)Scheme 1. Synthetic Approaches Towards the Synthesis of Formula (I) Compounds

[0175] Compounds of Formula (I) can be prepared using processes as illustrated in Scheme 1. Compounds of formula I-A are subjected to a palladium cross-coupling reaction with pyrazole boronic acids or esters to generate compounds I-B that are further reacted with an alchoxycarbonyl isothiocyanate, followed by reaction with hydroxylammonium chloride in the presence of a base like diisopropylethylamine (DIEA) to generate compounds of formula I-C. Alternatively, compounds I-A can first be reacted with an alchoxycarbonyl isothiocyanate, followed by reaction with hydroxylammonium chloride in the presence of a base like di isopropyl ethyl amine (DIEA) to generate a compounds of formula I-D that, in turn, can be subjected to a palladium cross-coupling reaction with pyrazole boronic acids or esters to generate compounds of formula I-C. Sandmeyer reaction with compounds I-C affords compounds I-E that can be subjected to a nucleophilic aromatic substitution (e.g., S\Ar) or a number of cross-coupling reactions, including Buchwald-Hartwig amination and copper-catalyzed aminations to give compounds of formula I-F. Alternatively, compounds of formula I-C can undergo reductive amination with corresponding ketones or aldehydes to generate directly compounds of formula I- F. Final protecting group deprotection of the pyrazolo group affords compounds of formula (I). Atany step of the synthesis, when R2 is halogen or hydroxy or amine, the compounds can be transformed to alternative R2 groups by various reactions, including S\Ar, O-alkylation, metal cross coupling reactions, reductive amination, etc.Scheme 2. Alternative Synthetic Approaches Towards the Synthesis of Formula (I) Compounds

[0176] Compounds of Formula (I) can also be prepared using processes as illustrated in Scheme 2. Compounds of formula I-D are subjected to a Sandmeyer reaction to generate compounds I-G, that are further reacted with a palladium cross-coupling reaction with pyrazole boronic acids or esters to generate compounds I-E. Alternatively, compounds of formula I-D can undergo reductive amination to form compounds I-H and then be subjected to a palladium crosscoupling reaction with pyrazole boronic acids or esters to generate compounds of formula I-F. Furthermore, compounds of formula I-H can first be converted to the boronic esters or acids of formula I-I and then be subjected to a palladium cross-coupling reaction with pyrazole halides to form compounds of formula I-F. In turn, compounds I-E can be subjected to a nucleophilic aromatic substitution (e.g., S\ Ar) or a number of cross-coupling reactions, including Buchwald- Hartwig amination and copper-catalyzed aminations to give compounds of formula I-F, which after final protecting group deprotection of the pyrazolo group affords compounds of formula (I). At any step of the synthesis, when R2 is halogen or hydroxy or amine, the compounds can betransformed to alternative R2 groups by various reactions, including S\Ar, O-alkylation, metal cross coupling reactions, reductive amination, etc.Scheme 3. Synthetic Approaches Towards the Synthesis of Formula (II) Compounds

[0177] As highlighted in Scheme 3, similar reaction sequences as described in Scheme 1 can also be performed for the synthesis of compounds of Formula (II).

[0178] Analytical methods for LC-MS analysis were performed using the noted columns unless described otherwise in the experimental procedures:

[0179] Analytical method 1: 10% B to 0.01 min, then 10-90% B to 2.00 min, hold at 90% B to 2.80 min and return to 10% B at 2.88 min, where A is Water / 5mM NH4HCO3 and B is ACN. Column: Shim-pack Scepter Cl 8.

[0180] Analytical method 2: 10% B to 0.01 min, then 10-70% B to 1.9 min, then 70- 90% B to 2 min, hold at 90% B to 2.7 min and return to 10% B at 2.75 min, where A is Water / 5mM NH4HCO3 and B is ACN. Column: Shim-Pack scepterCI 8.

[0181] Analytical method 3: 20% B to 0.01 min, then 20-70% B to 1.9 min, then 70- 90% B to 2 min, hold at 90% B to 2.7 min and return to 20% B at 2.75 min, where A is Water / 5mM NH4HCO3 and B is ACN. Column: Shim-Pack scepterCI 8.

[0182] Analytical method 4: 10% B to 0.01 min, then 10%-60% B to 1 .9 min, then 60%-90% B to 2 min, hold at 90% B to 2.7 min and return to 10% B at 2.75 min, where A is Water / 5mM NH4HCO3 and B is ACN. Column: Shim-Pack Velox SP-C18.

[0183] Analytical method 7: 10% B to 0.01 min, then 10-95% B to 2.00 min, hold at 95% B to 2.80 min and return to 10% B at 2.88 min, where A is Water / 5mM NH4HCO3 and B is ACN. Column: Shim-Pack scepterCI 8.

[0184] Analytical method 10: 5% B to 0.01 min, then 5-100% B to 2.00 min, hold at 100% B to 2.70 min and return to 5% B at 2.75 min, where A is Water / 0.09%FA and B is ACN / 0.1%FA. Column: Luna Omega PS Cl 8.

[0185] Analytical method 11: 5% B to 0.01 min, then 5-40% B to 1.90 min, then 40- 100% B to 2.00 min, hold at 100% B to 2.70 min and return to 5% B at 2.75 min, where A is Water / 0.09%FA and B is ACN / 0.1%FA. Column: Luna Omega PS C18.

[0186] Analytical method 12: 5% B to 0.01 min, then 5-50% B to 1.80 min, then 50- 100% B to 2.20 min, hold at 100% B to 2.70 min and return to 5% B at 2.75 min, where A is Water / 0.09%FA and B is ACN / 0.1%FA. Column: Luna Omega PS C18.

[0187] Analytical method 13: 5% B to 0.01 min, then 5-60% B to 1.70 min, then 60- 100% B to 2.20 min, hold at 100% B to 2.70 min and return to 5% B at 2.75 min, where A is Water / 0.09%FA and B is ACN / 0.1%FA. Column: Luna Omega PS C18.

[0188] Analytical method 15: 10% B to 0.01 min, then 10-60% B to 1.70 min, then 60- 100% B to 2.20 min, hold at 100% B to 2.70 min and return to 5% B at 2.75 min, where A is Water / 0.09%FA and B is ACN / 0.1%FA. Column: Luna Omega PS C18.

[0189] Analytical method 17: 0% B to 0.01 min, then 0-90% B to 2.0 min, hold at 90% B to 2.8 min and return to 0% B at 2.88 min, where A is 6.5mM NH4HCO3 in Water / Acetonitrile (9: l,v / v), pH=10 and B is ACN. Column: Kinetex EVO C18 -100A.

[0190] Analytical method 18: 0% B to 0.01 min, then 0-55% B to 1.9 min, then 55-90% B to 2 min, hold at 90% B to 2.7 min and return to 0% B at 2.75 min, where A is 6.5mM NH4HCO3 in Water / Acetonitrile (9:l,v / v), pH=10 and B is ACN. Column: Kinetex EVO C18 - 100A.

[0191] Analytical method 23: 0% B to 0.01 min, then 0-40% B to 1.2 min, then 40-90% B to 3 min, hold at 90% B to 2.7 min and return to 0% B at 2.75 min, where A is 6.5mMNH4HCO3 in Water / Acetonitrile (9:l,v / v), pH=10 and B is ACN. Column: Kinetex EVO Cl 8 - 100A.

[0192] Analytical method 25: 0% B to 0.01 min, then 0-50% B to 1.9 min, then 50-90% B to 2 min, hold at 90% B to 2.7 min and return to 0% B at 2.75 min, where A is 6.5mM NH4HCO3 in Water / Acetonitrile (9:l,v / v), pH=10 and B is ACN. Column: Kinetex EVO C18 - 100A.

[0193] Analytical method 26: 5% B to 0.01 min, then 5-100% B to 2.00 min, hold at 100% B to 2.70 min and return to 5% B at 2.75 min, where A is Water / 0.05%TFA and B is ACN / 0.05%TFA. Column: XSelect HSS T3.

[0194] Analytical method 27: 5% B to 0.01 min, then 5-50% B to 1.90 min, then 60- 100% B to 2.00 min, hold at 100% B to 2.70 min and return to 5% B at 2.75 min, where A is Water / 0.05%TFA and B is ACN / 0.05%TFA. Column: XSelect HSS T3.

[0195] Analytical method 28: 5% B to 0.01 min, then 5-100% B to 1.10 min, hold at 100% B to 1.75 min and return to 5% B at 1.8 min, where A is Water / 0.05%TFA and B is ACN / 0.05%TFA. Column: XSelect HSS T3.

[0196] Analytical method 29: 5% B to 0.01 min, then 5-40% B to 1.80 min, then 40- 100% B to 2.00 min, hold at 100% B to 2.70 min and return to 5% B at 2.75 min, where A is Water / 0.05%TFA and B is ACN / 0.05%TFA. Column: XSelect HSS T3.

[0197] Analytical method 30: 5% B to 0.01 min, then 5-60% B to 1.90 min, then 60- 100% B to 2.00 min, hold at 100% B to 2.70 min and return to 5% B at 2.75 min, where A is Water / 0.05%TFA and B is ACN / 0.05%TFA. Column: XSelect HSS T3.

[0198] Analytical method 33: 5% B to 0.01 min, then 5-100% B to 2.00 min, hold at 100% B to 2.70 min and return to 5% B at 2.75 min, where A is Water / 0.05%TFA and B is ACN / 0.05%TFA. Column: HALO C18.

[0199] Analytical method 49: 10% B to 0.01 min, then 10-40% B to 2.00 min, then 40- 100% B to 2.10 min, hold at 100% B to 2.80 min and return to 5% B at 2.85 min, where A is Water / 0.1%FA and B is ACN / 0.07%FA. Column: Ultimate XB-C18.

[0200] Analytical method 62: 10% B to 0.01 min, then 10-90% B to l.Omin, hold at 90% B to 1.50min and return to 10% B at 1.51 min, where A is 5mM NH4HCO3 in Water and B is ACN. Column: Kinetex EVO.

[0201] Analytical method 64: 0% B to 0.01 min, then 10-90% B to 2.0 min, hold at 90% B to 2.8min and return to 10% B at 2.88 min, where A is 6.5mM NH4HCO3 in Water, pH=10 and B is ACN. Column: Shim Pack Scepter C18.

[0202] Analytical method 65: 0% B to 0.01 min, then 0-90% B to 1.20 min, hold at 90% B to 1.80 min and return to 0% B at 1.82 min, where A is 6.5mM NH4HCO3 inWater / Acetonitrile(9: l,v / v), pH=10 and B is ACN. Column: Kinetex EVO C18 -100A.

[0203] Analytical method 66: 5% B to 0.01 min, then 5-100% B to 1.00 min, hold at 100% B to 1.40 min and return to 5% B at 1.42 min, where A is Water / 0.05% TFA and B is ACN / 0.5%TFA. Column: Shim-pack.

[0204] Analytical method 67: 5% B to 0.01 min, then 5-100% B to 1.20 min, hold at 100% B to 1.80 min and return to 5% B at 1.82 min, where A is Water / 0.09%FA and B is ACN / 0.1%FA. Column: Luna Omega PS Cl 8.

[0205] Analytical method 68: 5% B to 0.01 min, then 5-100% B to 1.00 min, hold at 100% B to 1.40 min and return to 5% B at 1.42 min, where A is Water / 0.1% FA and B is ACN / 0.1%FA. Column: HALO.

[0206] Analytical method 69: 10% B to 0.01 min, then 10-90% B to 1.20 min, hold at 90% B to 1.80 min and return to 10% B at 1.82 min, where A is Water / 5mM NH4HCO3 and B is ACN. Column: Shim-Pack scepter C18.

[0207] Analytical method 70: 5% B to 0.01 min, then 5-100% B to 0.70 min, hold at 100% B to 1.10 min and return to 5% B at 1.42 min, where A is Water / 0.05% TFA and B is ACN / 0.5%TFA. Column: Shim-pack.

[0208] Analytical method 72: 10% B to 0.01 min, then 10-95% B to 0.9 min, hold at 95% B to 1.30 min and return to 10% B at 1.33 min, where A is 5mM NH4HCO3 in Water and B is ACN. Column: Shim-Pack scepter Cl 8.

[0209] Analytical method 73: 5% B to 0.01 min, then 5-95% B to 1.00 min, hold at 95% B to 1.50 min and return to 5% B at 1.51 min, where A is 5mM NH4HCO3 in Water and B is ACN. Column: Shim-Pack scepter C18-120.

[0210] Analytical method 74: 5% B to 0.01 min, then 5-100% B to 1.00 min, hold at 100% B to 1.40 min and return to 5% B at 1.42 min, where A is Water / 0.05% TFA and B is ACN / 0.5%TFA). Column: HALOWA column.

[0211] Analytical method 82: 5% B to 0.01 min, then 5%-100% B to 2.00 min, hold at 100% B to 2.70 min and return to 5% B at 2.75 min, Where A is Water / 0.05%TFA and B is ACN / 0.05%TFA. Column: Ascentis Express Cl 8.

[0212] Analytical method 83: 5% B to 0.01 min, then 5-50% B to 1.90 min, then 50- 100% B to 2.00 min, hold at 100% B to 2.70 min and return to 5% B at 2.75 min, where A is Water / 0.05%TFA and B is ACN / 0.05%TFA. Column: XSelect HSS T3.

[0213] Analytical method 84:10% B to 0.01 min, then 10-95% B to 2.00 min, hold at 95% B to 2.80 min and return to 10% B at 2.75 min, where A is Water / 5mM NH4HCO3 and B is ACN. Column: Shim-Pack Scepter.

[0214] Analytical method 85: 0% B to 0.01 min, then 0-35% B to 1.9 min, then 35-90% B to 2 min, hold at 90% B to 2.7 min and return to 0% B at 2.75 min, where A is 6.5mM NH4HCO3 in Water / Acetonitrile (9:l,v / v), pH=10 and B is ACN. Column: Kinetex EVO C18 - 100A.

[0215] Analytical method 86: 0% B to 0.01 min, then 0-45% B to 1.9 min, then 45-90% B to 2.0 min, hold at 90% B to 2.7 min and return to 0% B at 2.75 min, where A is 6.5 mM NH4HCO3 in Water / ACN (9: l,v / v), pH=10 and B is ACN. Column: Kinetex EVO C18 -100A

[0216] Analytical method 87: 0% B to 0.01 min, then 0-90% B to 1.20 min, hold at 90% B to 1.80 min and return to 0% B at 1.82 min, where A is Water / 5mM NH4HCO3 and B is ACN. Column: Shim-Pack Cl 8.

[0217] Analytical method 88: 10% B to 0.01 min, then 10%-60% B to 1.9 min, then 60%-95% B to 2 min, hold at 95% B to 2.7 min and return to 10% B at 2.75 min, where A is Water / 5mM NH4HCO3 and B is ACN. Column: Shim-Pack scepterCI 8.

[0218] Analytical method 89: 0% B to 0.01 min, then 0-40% B to 1.9 min, then 40-90% B to 2 min, hold at 90% B to 2.7 min and return to 0% B at 2.75 min, where A is 6.5mM NH4HCO3 in Water / Acetonitrile (9:l,v / v), pH=10 and B is ACN. Column: Kinetex EVO C18 - 100A.

[0219] Analytical method 90: 10% B to 0.01 min, then 10%-60% B to 1.9 min, then 60%-90% B to 2 min, hold at 90% B to 2.7 min and return to 10% B at 2.75 min, where A is Water / 5mM NH4HCO3 and B is ACN. Column: Shim-Pack scepterCI 8.

[0220] Analytical method 91: 10% B to 0.01 min, then 10-50% B to 1.9 min, then 50- 90% B to 2.0 min, hold at 90% B to 2.7 min and return to 10% B at 2.75 min, where A is Water / 5mM NH4HCO3 and B is ACN. Column: Shim-pack Scepter Cl 8.

[0221] Analytical method 93: 5% B to 0.01 min, then 5-95% B to l.Omin, hold at 95% B to 1.50min and return to 5% B at 1.51 min, where A is 5mM NH4HCO3 in Water and B is ACN. Column: Kinetex EVO C18 -100A

[0222] Analytical method 96: 0% B to 0.01 min, then 0-100% B to 2.00 min, hold at 100% B to 2.70 min and return to 0% B at 2.75 min, where A is Water / 0.05%TFA and B is ACN / 0.05%TFA. Column: XSelect HSS T3.Example 2 - Synthesis of IntermediatesSynthesis of 5-Chloro-6-( 1 -( 1 -ethoxy ethyl)- lH-pyrazol-4-yl)-[ 1 ,2,4]triazolo[ 1 ,5-a]pyrazin-2-

[0223] Step 1 : Into a 2 L 3 -necks flask purged and maintained with an inert atmosphere of nitrogen was placed a solution of 5-bromo-6-chloropyrazin-2-amine (60.0 g, 287.853 mmol, 1.0 equiv) and l-(l-ethoxyethyl)-4-(4,4,5,5-tetramethyl-l,3,2-dioxaborolan-2-yl) pyrazole (76.6g, 287.853 mmol, 1.0 equiv) in dioxane (1.2 L). Water (240.0 mb), K3PO4 (122.2 g, 575.706 mmol, 2.0 equiv), and Pd(dppf)Ch CH2CI2 (11.7 g, 14.39 mmol, 0.05 equiv) were added to the mixture at room temperature. The mixture was bubbled with N2 for 1 min. The mixture was then stirred overnight at 90 °C. The reaction was cooled to room temperature. The resulting solution wasdiluted with 1 L of water and extracted with DCM (2 x 800 mL). The organic layers were combined, washed with brine, dried with Na2SO4, and fdtered. The filtrate was concentrated and purified by column chromatography (60% EA in PE) to afford 6-chloro-5-(l-(l-ethoxyethyl)-lH-pyrazol-4- yl)pyrazin-2-amine (Int-la, 60.0 g, 77 % yield) as an off-white solid. LCMS (ESI) calculated for C11H14CIN5O (M+H)+: 268.15, found: 196.05, RT=0.964 min (analytical method 62).

[0224] Step 2 : Into a 2 L 3 -necks flask purged and maintained with an inert atmosphere of nitrogen was placed a solution of Int-la (58.0 g, 216.64 mmol, 1.0 equiv) in ACN (1.2 L) and treated at room temperature with ethyl N-carbothioylcarbamate (42.6 g, 324.966 mmol, 1.5 equiv). The mixture was bubbled with N2 for 1 min. The mixture was then stirred for 2 hours at 60 °C. The reaction was cooled to room temperature. The resulting mixture was concentrated under reduced pressure to afford crude ethyl N-({6-chloro-5-[l-(l-ethoxyethyl)-lH-pyrazol-4-yl] pyrazin-2-yl} carbarn othioyl)carbamate (Int-lb, 91.0 g) as a yellow solid. The crude product was used in the next step directly without further purification. LCMS (ESI) calculated for C15H19CIN6O3S (M+H)1: 399.20, found: 399.20, RT=0.959 min (Shim-pack Scepter C18-120 column, analytical method 62).

[0225] Step 3 : Into a 2 L 3 -necks flask purged and maintained with an inert atmosphere of nitrogen was placed a solution of Int-lb (91.0 g, 205.33 mmol, 1.0 equiv, 90%) in MeOH (900.0 mL) and EtOH (900.0 mL), hydroxylamine hydrochloride (42.8 g, 615.990 mmol, 3.0 equiv), and DIEA (79.6 g, 615.990 mmol, 3.0 equiv) were added to the mixture at room temperature. The mixture was bubbled with N2 for 1 min. The mixture was then stirred for 2 hours at 900C. The reaction was cooled to room temperature. The resulting solution was diluted with 2 L of water and extracted with DCM (2 x 700 mL). The organic layers were combined, washed with brine, dried with Na2SO4, filtered, and evaporated to dryness under reduced pressure to afford 5-chloro-6-(l- (l-ethoxyethyl)-lH-pyrazol-4-yl)-[l,2,4]triazolo[l,5-a]pyrazin-2-amine (Int-1, 85.0 g) as a yellow solid. The crude product was used in the next step directly without further purification. LCMS (ESI) calculated for C12H14CIN7O (M+H)+: 308.10, found: 308.10, RT=1.051 min (analytical method 64).Synthesis of 2-Bromo-5-chloro-6-(l-(l -ethoxy ethyl)- lH-pyrazol-4-yl)-[ 1,2, 4]triazolo[ 1,5- a]pyrazine (Int-2)lnt-1 lnt-2

[0226] Into a 1 L 3-necks flask purged and maintained with an inert atmosphere of nitrogen was placed a solution of CuBn (14.3 g, 64.340 mmol, 1.2 equiv) in ACN (500.0 mL), and then tBuNCh (14.7 g, 128.681 mmol, 2.4 equiv, 90%) and Int-1 (25.0 g, 53.617 mmol, 1.0 equiv, 66%) were added at room temperature. The mixture was bubbled with N2 for 1 min. The mixture was stirred for 30 min at room temperature. The resulting mixture was filtered. The filter cake was washed with ACN (2 x 20 mL). The filtrate was concentrated under reduced pressure and purified by column chromatography (40% EA in PE) to afford 2-bromo-5-chloro-6-(l-(l-ethoxyethyl)-lH- pyrazol-4-yl)-[l,2,4]triazolo[l,5-a]pyrazine (Int-2, 9.0 g, 45 % yield) as a brown solid. LCMS (ESI) calculated for C12H12BrClN6O (M+H)1: 372.62, found: 373.00, RT=1.271 min (analytical method 17); 'H-NMR (400 MHz, DMSO-d6,ppm}. 6 931 (s, 1H), 8.66 (d, J= 0.8 Hz, 1H), 8.23 (s, 1H), 5.69 (q, J= 5.9 Hz, 1H), 3.49 (dq, J = 9.5, 7.0 Hz, 1H), 3.26 (dq, J = 9.6, 7.0 Hz, 1H), 1.67 (d, J= 6.0 Hz, 3H), 1.25 - 1.15 (m, 1H), 1.07 (t, J = 7.0 Hz, 3H).Synthesis of 4-[2-Bromo-5-(2,2,2-trifluoroethoxy)-[l,2,4]triazolo[l,5-a]pyrazin-6-yl]-l-(l- ethoxy ethyl)pyrazole (Int-3)Int-2

[0227] Into a 40 ml vial purged and maintained with an inert atmosphere of nitrogen was placed a solution of Int-2 (500.0 mg, 1.345 mmol, 1.0 equiv) in dioxane (10 mL) and 2,2,2- trifluoroethan-l-ol (134.6 mg, 1.345 mmol, 1.0 equiv) was added. The mixture was cooled to 0°C and treated with NaH (32.3 mg, 1.345 mmol, 1.0 equiv). The mixture was stirred for 5 min at 0 °C and then for 2 hours at 120 °C. The reaction was cooled to room temperature. The resulting solution was diluted with 10 mL of water and extracted with ethyl acetate (3 x 10 mL). The organic layerswere combined, washed with brine, dried with NazSCh, filtered, and concentrated under reduced pressure. The crude was purified by reverse phase flash chromatography [column, Cl 8; Mobile Phase A: Water (0.1% NH4HCO3), Mobile Phase B: ACN; Flow rate: 60 mL / min; Gradient: 40% B to 80% B in 30 min; Wave Length: 210 nm; RT1 (min): 18] to afford 4-[2-bromo-5-(2,2,2- trifluoroethoxy)-[l,2,4]triazolo[l,5-a]pyrazin-6-yl]-l-(l-ethoxyethyl)pyrazole (Int-3, 200 mg, 34% yield) as a white solid. LCMS (ESI) calculated for C14H14BrF3N6O2 (M+H)+: 435.03, found: 437.05, RT=1.003 min (analytical method 28); 'H-NMR (400 MHz, DMSO-de, ppm): 8 9.24 (s, 1H), 8.36 (s, 1H), 8.06 (s, 1H), 5.67 (q, J = 5.9 Hz, 1H), 5.31 (qd, J = 8.9, 1.4 Hz, 2H), 3.48 (dq, J = 9.5, 7.0 Hz, 1H), 3.29 - 3.15 (m, 1H), 1.64 (d, J = 6.0 Hz, 3H), 1.07 (t, J = 7.0 Hz, 3H);19F NMR (376 MHz, DMSO-d6, ppm) 8 -72.78.Synthesis of 6-(l-(l-Ethoxyethyl)-lH-pyrazol-4-yl)-5-(2,2,2-trifluoroethoxy)-[l,2,4]triazolo[l,5- a]pyrazin-2-amine (Int-4)lnt-1 lnt-4

[0228] Into a 40 ml vial purged and maintained with an inert atmosphere of nitrogen was placed a solution of Int-1 (1.0 g, 3.249 mmol, 1.0 equiv) in dioxane (10 mL). 2,2,2-Trifluoroethan- l-ol (325.0 mg, 3.249 mmol, 1.0 equiv) was added and the mixture was cooled to 0°C. Then NaH (78.0 mg, 3.249 mmol, 1.0 equiv) was added and the mixture was stirred for 5 min at 0°C and 2 hours at 120 °C. The reaction was cooled to room temperature. The resulting solution was diluted with 10 mL of water and extracted with ethyl acetate (3 x 10 mL). The organic layers were combined, washed with brine, dried with Na?SO4, filtered, and concentrated under reduced pressure to afford 6-(l-(l-ethoxyethyl)-lH-pyrazol-4-yl)-5-(2,2,2-trifluoroethoxy)- [l,2,4]triazolo[l,5-a]pyrazin-2-amine (Int-4, 600 mg, 50% yield) as an off-white solid. The crude was used in the next reaction without further purification. LCMS (ESI) calculated for C14H16F3N7O2 (M+H)+: 372.13, found:372.20. RT=1.024 min (analytical method 62).Synthesis of 7-Chloro-8-(2,2,2-trifluoroethoxy)-[l,2,4]triazolo[l,5-c]pyrimidin-2-amine (Int-5)Y=34% lnt-5bStep 3 lnt-5

[0229] Step 1 : Into a 500 mL 3-necked round-bottom flask at room temperature were added 4-amino-6-chloropyrimidin-5-ol (40.0 g, 274.820 mmol, 1.0 equiv) and 2,2,2-trifluoroethyl trifluoromethanesulfonate (70.2 g, 302.302 mmol, 1.1 equiv) in DMF (200.0 mL), followed by CS2CO3 (107.45 g, 329.784 mmol, 1.2 equiv). The resulting mixture was stirred overnight at room temperature and extracted with EA (3 x 200 mL). The combined organic layers were washed with brine (3 x 1000 mL), dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure to afford 6-chloro-5-(2,2,2-trifluoroethoxy) pyrimidin-4-amine (Int-5a, 53.0 g, crude) as a yellow solid. The crude was used in the next step without further purification.

[0230] Step 2 : Into a 40mL vial at room temperature were added Int-5a (4.5 g, 19.774 mmol, 1.0 equiv) and ACN (9.0 mL), followed by ethyl N-carbothioylcarbamate (3.9 g, 29.661 mmol, 1.5 equiv). The resulting mixture was stirred overnight at 80°C. The mixture was then allowed to cool down to room temperature and the reaction progress was confirmed by LC-MS. The mixture was concentrated under reduced pressure to afford ethyl N-{[6-chloro-5-(2,2,2- tri fluoroethoxy) pyrimidin-4-yl] carbamothioyl} carbamate (Int-5b, 7.0 g, crude) as a brown oil that was used in the next step directly without further purification. LCMS (ESI) calculated for C10H10CIF3N4O3S (M)+: 358.01, found: 358.70. RT=0.699 min (analytical method 68).

[0231] Step 3 : Into a 250mL 3-necked round-bottom flask at room temperature were added Int-5b (7.0 g, 19.514 mmol, 1.0 equiv) and EtOH (70.0 mL), followed by hydroxylamine hydrochloride (4.07 g, 58.542 mmol, 3.0 equiv). The resulting mixture was stirred for 3 hours at60°C and reaction progress confirmed by LC-MS. The mixture was then allowed to cool down to room temperature and concentrated under reduced pressure. The residue was purified by reverse phase flash with the following conditions (Reversal phase Combi-Flash; Column: WelFlash Cl 8- I Column, Regular Cl 8 20-40qm, 330g; Condition: Mobile Phase A: Water (10 mmol / L NH4HCO3), Mobile Phase B: ACN; Flow rate: 100 mL / min; Gradient: 15% B to 45% B in 30min; Wave Length: 254nm / 210nm; RTl(min): 28.0) to afford 7-chloro-8-(2,2,2-trifluoroethoxy)- [1,2,4] triazolo[l,5-c] pyrimidin-2-amine (Int-5, 1.8 g, 34%) as a yellow solid. LCMS (ESI) calculated for C7H5QF3N5O (M+H)+: 268.01, found: 267.95. RT=0.768 min (analytical method 66).Synthesis of 7-(l-(l-Ethoxyethyl)-lH-pyrazol-4-yl)-8-(2,2,2-trifluoroethoxy)-[l,2,4]triazolo[l,5- c]pyrimidin-2-amine (Int-6)-

[0232] Step 1 : Into a 2 L 3-necked round-bottom flask were added at room temperature Int-5a (53 g, 232.895 mmol, 1 equiv) and l-(l-ethoxyethyl)-4-(4,4,5,5-tetramethyl-l,3,2- dioxaborolan-2-yl)pyrazole (74.38 g, 279.474 mmol, 1.2 equiv) in dioxane (1060 mL) and H2O (212 mL). The mixture was then treated under nitrogen atmosphere with K3PO4 (98.87 g, 465.790 mmol, 2 equiv) and Pd(dppf)C12 (17.04 g, 23.290 mmol, 0.1 equiv). The resulting mixture was stirred overnight at 100°C. The mixture was allowed to cool down to room temperature and was filtered. The filter cake was washed with 1,4-di oxane (3 x 50 mL). The filtrate was extracted with EA (3 x 250 mL). The combined organic layers were washed with brine (3 x 250 mL), dried overanhydrous Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography, eluted with PE / EA (1 :5) to afford 6-( 1-(1 -ethoxy ethyl)- 1H- pyrazol-4-yl)-5-(2,2,2-trifluoroethoxy)pyrimidin-4-amine (Int-6a, 71 g, 75% for 2 steps) as a yellow oil. LCMS (ESI) calculated for C13H16F3N5O2 (M+H)+: 332.13, found: 332.10; RT= 0.878 min (HALOWA column, analytical method 66); 'H-NMR (400 MHz, DMSO-t / c, ppm). 8 8.35 (s, 1H), 8.17 (s, 1H), 8.03 (s, 1H), 6.92 - 6.73 (m, 2H), 5.69 - 5.55 (m, 1H), 4.59 - 4.43 (m, 2H), 3.51 - 3.40 (m, 1H), 3.30 - 3.16 (m, 1H), 1.61 (d, J= 6.0 Hz, 3H), 1.11 - 0.99 (m, 3H).

[0233] Step 2 : Into a 3 L three-necked flask were placed at room temperature a solution of Int-6a (70 g, 211.4 mmol, 1 equiv) in ACN (1400 mL). Ethyl N-carbothioylcarbamate (42 g, 317.1 mmol, 1.5 equiv) was added and the mixture was stirred overnight at 60°C. The mixture was then concentrated under vacuum to afford ethyl N-({6-[l-(l-ethoxyethyl)-lH-pyrazol-4-yl]-5- (2,2,2-trifluoroethoxy)pyrimidin-4-yl}carbamothioyl) carbamate (Int-6b, 97.5 g, crude) as a brown oil. The crude was used in the next step without further purification LCMS (ESI) calculated for C17H21F3N6O4S (M+H)1: 463.13 , found: 463.20; RT= 1.119 min (analytical method 65).

[0234] Step 3 : Into a 3 L three-necked flask were placed at room temperature a solution of Int-6b (70 g, 151.41 mmol, 1 equiv) in MeOH (700 mL) and EtOH (700 mL). Hydroxylamine hydrochloride (31.6 g, 454.7 mmol, 3 equiv) and DIEA (58.6 g, 454.7 mmol, 3 equiv) were added. The mixture was then stirred for 1 hour at 60°C, concentrated under vacuum and extracted with EA (350mL) and washed with H2O (400mL). The organic layer was concentrated under vacuum and the resulting residue was purified by Combi-Flash with the following conditions (Column: Cis; Mobile Phase A: Water (0.1% FA), Mobile Phase B: ACN; Flow rate: 100 mL / min; Gradient: 15% B to 50% B in 25 min; Wave Length: 254 / 210 nm; RTl(min): 20) to afford 7-(l-(l -ethoxy ethyl)- lH-pyrazol-4-yl)-8-(2,2,2-trifluoroethoxy)-[l,2,4]triazolo[l,5-c]pyrimidin-2-amine (Int-6, 27.3 g, 48% for 2 steps) as a grey solid. LCMS (ESI) calculated for C14H16F3N7O6 (M+H)+: 372.13, found: 372. 10; RT= 1.090 min (HALOWA column, analytical method 66); 'H-NMR (400 MHz, DMSO- d6,ppm) 8 9.15 (s, 1H), 8.39 (s, 1H), 8.10 (s, 1H), 6.62 - 6.41 (m, 2H), 5.68 - 5.60 (m, 1H), 5.40 - 5.30 (m, 2H), 3.50 - 3.43 (m, 1H), 3.28 - 3.21 (m, 1H), 1.63 (d, 3H), 1.09 - 1.04 (m, 3H).Synthesis of 2-Bromo-7-(l-(l -ethoxyethyl)- lH-pyrazol-4-yl)-8-(2, 2, 2-trifluoroethoxy)-[ 1 ,2,4]triazolo[ 1 ,5-c]pyrimidine (Int-7)lnt-6 lnt-7

[0235] Into a 1 L 3 -necked round-bottom flask were added at room temperature tert-butyl nitrite (12 g, 116.442 mmol, 2.4 equiv) and CuBn (13 g, 58.221 mmol, 1.2 equiv) in ACN (180 mL). The mixture was then treated with a solution of Int-6 (18 g, 48.517 mmol, 1 equiv) in ACN (180 mL) and stirred for 0.5 hour at room temperature. The mixture was filtered and the filtrate was purified by combi-flash (ACN: H2O with 0.1% FA 50%~100%, 30 min) to afford 4-[2-bromo- 8-(2,2,2-trifluoroethoxy)-[ 1 ,2,4] tri azolof 1 ,5-c]pyrimidin-7-yl]- 1 -(1 -ethoxy ethyl) pyrazole (Int-7, 14.5 g, 69% yield) as a white solid. LCMS (ESI) calculated for CuHuBrFsNeCh (M+H)+: 435.03, found:435.05; RT 1.054 min (HALOWA column, analytical method 65); 'H-NMR (400 MHz, DMSO-tL. ppm)'. 8 9.60 (s, 1H), 8.47 (s, 1H), 8.16 (s, 1H), 5.73 - 5.62 (m, 1H), 5.45 - 5.31 (m, 2H), 3.56 - 3.42 (m, 1H), 3.32 - 3.20 (m, 1H), 1.63 (d, J= 5.9 Hz, 3H), 1.12 - 1.00 (m, 3H).Synthesis of 8-Chloro-7-(l-(l -ethoxy ethyl )-lH-pyrazol-4-yl)-[ 1,2, 4]triazolo[l,5-c]pyrimidin-2-

[0236] Step 1 : Into a 3 L 3-necked round bottle purged and maintained with an inert atmosphere of nitrogen was placed a solution of 5,6-dichloropyrimidin-4-amine (130.0 g, 792.731 mmol, 1.0 equiv), l-(l-ethoxyethyl)-4-(4,4,5,5-tetramethyl-l,3,2-dioxaborolan-2-yl)pyrazole (232.1 g, 872.004 mmol, 1.1 equiv), and Na2C0.3 (252.06 g, 2378.193 mmol, 3.0 equiv) in dioxane (1300 mL) and H2O (325.0 mb). The reaction mixture was evacuated and flushed 3 times with nitrogen. Pd(dppf)C12.CH2C12 (45.20 g, 55.491 mmol, 0.07 equiv) was added and the mixture was stirred overnight at 90 °C under nitrogen. After confirming the reaction progress by LC-MS, the mixture was cooled to room temperature and poured into water (1000 ml). The resulting mixture was extracted with ethyl acetate (3 x 1000 mL). The combined organic lavers were washed with brine (500ml), dried over anhydrous Na2SO4, filtered, and the filtrate was concentrated under reduced pressure. The residue was purified by flash chromatography (0-70% EA:PE) to afford 5- chloro-6-(l-(l-ethoxyethyl)-lH-pyrazol-4-yl)pyrimidin-4-amine (Int-8a, 162.0 g, 76.3%) as a light-yellow solid. LCMS (ESI) calculated for C11H14CIN5O (M+H)+: 268.1, found: 268.1; RT=0.574 min (analytical method 28).

[0237] Step 2 : Into a 5 L 3-necked round bottle purged and maintained with an inert atmosphere of nitrogen was placed a solution of Int-8a (130.0 g, 485.582 mmol, 1.0 equiv) in toluene (2600.0 mL). Ethyl N-carbothioylcarbamate (127.4 g, 971.164 mmol, 2.0 equiv) was then added at room temperature. The mixture was stirred for 24 hours at 80 °C. The resulting mixture was concentrated under reduced pressure to afford ethyl N-({5-chloro-6-[l-(l-ethoxyethyl)-lH- pyrazol-4-yl]pyrimidin-4-yl}carbamothioyl)carbamate (Int-8b, 270.0 g, crude) as a brown oil. LCMS (ESI) calculated for C15H19CIN6O3S (M+H)+: 399.1, found: 399.3; RT 0.79 l min (analytical method 66).

[0238] Step 3 : Into a 5 L 3-necked round bottle purged and maintained with an inert atmosphere of nitrogen was placed a solution of Int-8b (270.0 g, 676.912 mmol, 1.0 equiv) in MeOH (1300.0 mL) and EtOH (1300.0 mL). Hydroxylamine hydrochloride (141.1 g, 2030.736 mmol, 3.0 equiv) and DIEA (262.5 g, 2030.736 mmol, 3.0 equiv) were then added at room temperature. The mixture was stirred for 1 hour at 60 °C. The reaction mixture was cooled to room temperature. Most of the solvents were removed under reduced pressure and the resulted precipitate was collected by filtration and washed with EtOH (1000.0 mL) to afford 8-chloro-7- (1 -(1 -ethoxy ethyl)- lH-pyrazol-4-yl)-[l, 2, 4]triazolo[l,5-c]pyrimidin-2-amine (Int-8, 90.0 g, 60 % yield over 2 steps) as a white solid. LCMS (ESI) calculated for C12H14CIN7O (M+H)+: 308.1,found: 308.1 ; RT=0.673 min (analytical method 28);1HNMR (400 MHz, DMSO-^6, ppm).' 5 9.28 (s, 1H), 8.68 (d, J= 0.8 Hz, 1H), 8.27 (s, 1H), 6.65 (s, 2H), 5.75 - 5.60 (m, 1H), 3.54 - 3.42 (m, 1H), 3.29 - 3.19 (m, 1H), 1.65 (d, J= 6.0 Hz, 3H), 1.13 - 1.00 (m, 3H).Synthesis of 2-Bromo-8-chloro-7-(l -(1 -ethoxyethyl)- lH-pyrazol-4-yl)-[ 1,2, 4]triazolo[ 1,5- c]pyrimidine (Int-9)

[0239] Into a 1 L 3 -necked round bottle purged and maintained with an inert atmosphere of nitrogen was placed a solution of CuBr2 (13.9 g, 62.390 mmol, 1.2 equiv) and tert-butyl nitrite (12.9 g, 124.781 mmol, 2.4 equiv) in ACN (160.0 mL). The mixture was stirred for 5 min at room temperature and a solution of Int-8 (16.0 g, 51.992 mmol, 1.0 equiv) in ACN (160.0 mL) was added. The mixture was stirred for 30 min at room temperature. The mixture was then filtered and the filtrate was purified by combi-flash with the following conditions: Column: Cl 8; Mobile Phase A: Water(0.1% FA), Mobile Phase B: ACN; Flow rate: 100 mL / min; Gradient: 15% B to 85% B in 30 min; Wave Length: 220 nm; RTl(min): 19.00 to afford 2-bromo-8-chloro-7-(l-(l- ethoxyethyl)-lH-pyrazol-4-yl)-[l,2,4]triazolo[l,5-c]pyrimidine (Int-9, 12.0 g, 62.1% yield) as a white solid. LCMS (ESI) calculated for C 12H12BrCIN6O (M+H)+: 372.95, found: 372.95; RT-0.953 min (analytical method 67); 'HNMR (400 MHz, DMSO-dd6,ppm): 5 9.72 (s, 1H), 8.80 (s, 1H), 8.34 (s, 1H), 5.81 - 5.58 (m, 1H), 3.58 - 3.43 (m, 1H), 3.31 - 3.16 (m, 1H), 1.67 (d, J = 5.9 Hz, 3H), 1.14 - 0.98 (m, 3H).Synthesis of 8-Chloro-7-(l-(l-ethoxyethyl)-lH-pyrazol-4-yl)-[l,2,4]triazolo[l,5-a]pyridin-2- amine (Int-10)

[0240] Step 1 : Into a 500 ml 3-necked round bottle was placed at room temperature a solution of 4-bromo-3-chloropyridin-2-amine (15.0 g, 72.303 mmol, 1.0 equiv) and 1-(1- ethoxyethyl)-4-(4,4,5,5-tetramethyl-l,3,2-dioxaborolan-2-yl)pyrazole (21.2 g, 79.533 mmol, 1.1 equiv) in 1,4-dioxane (300 mL) and H2O (60 mL). The mixture was then treated with K3PO4 (30.7 g, 144.606 mmol, 2.0 equiv) and bubbled with N2 for 2 min. Pd(dppf)ChCH2C12 (5.9 g, 7.230 mmol, 0.1 equiv) was then added and the mixture was stirred overnight at 90°C. The reaction was cooled to room temperature. The resulting solution was diluted with 100 ml of water and extracted with ethyl acetate (3 x 100 mL). The organic layers were combined, washed with brine, dried with Na2SO4, fdtered, and the filtrate was concentrated and purified by column chromatography (28% EA:PE) to afford 3 -chloro-4-(l-(l -ethoxy ethyl)- lH-pyrazol-4-yl)pyridin-2-amine (Int-lOa, 18 g, 93%) as a yellow solid. LCMS (ESI) calculated for C12H15CIN4O (M+H)+: 267.09, found:267.09. RT=0.503 min (analytical method 66).

[0241] Step 2: Into a 500 ml 3-necked round bottle was placed a solution of Int-lOa (18.0 g, 67.484 mmol, 1.0 equiv) and ethyl N-carbothioylcarbamate (13.3 g, 101.226 mmol, 1.5 equiv) in ACN (360 mL). The mixture was stirred for 2 hours at 60°C. The reaction was cooled to room temperature and concentrated under reduced pressure to afford ethyl N-({3-chloro-4-[l-(l- ethoxyethyl)pyrazol-4-yl]pyridin-2-yl}carbamothioyl)carbamate (Int-lOb, 32.0 g, crude) as a brown solid. The crude product was used in the next step without further purification. LCMS (ESI) calculated for C16H20CIN5O3S (M+H)+: 398.10, found:398.10. RT=0.750 min (analytical method 66).

[0242] Step 3: Into a 500 ml 3-necked round bottle was placed a solution of Int-lOb (32.0 g, 64.341 mmol, 1.0 equiv) in MeOH (250 mL) and EtOH (250 mL). NH2OH.HCI (13.4 g, 193.023 mmol, 3.0 equiv) and DIEA (25 g, 193.023 mmol, 3.0 equiv) were then added to the mixture at room temperature and the mixture was stirred for 2 hours at 60°C. The reaction was cooled to room temperature. The resulting solution was diluted with NH4CI (100 ml) and extracted with DCM (3 x 100 mL). The organic layers were combined, washed with brine, dried with Na2SO4, filtered and concentrated to afford 8-chloro-7-(l-(l-ethoxyethyl)-lH-pyrazol-4-yl)- [l,2,4]triazolo[l,5-a]pyridin-2-amine (Int-10, 30.0 g, crude) as a green solid. The crude was used in the next step without further purification. LCMS (ESI) calculated for C13H15ClN6O (M+H)+: 307.10, found:307.10. RT=0.914 min (analytical method 62).Synthesis of 2-Bromo-8-chloro-7-(l -(1 -ethoxy ethyl)- lH-pyrazol-4-yl)-[ 1,2, 4]triazolo[ 1,5- a]pyridine (Int-11)Y=59%Int-10 Int-11

[0243] Into a 250 ml round bottle was placed a solution of CuBr? (1.8 g, 8.150 mmol, 1.0 equiv) and tert-butyl nitrite (1.7 g, 16.300 mmol, 2.0 equiv) in ACN (100 mL). The mixture was cooled to 0°C, treated with Int-10 (5 g, 8.150 mmol, 1.0 equiv) and then stirred for 0.5 hours at room temperature. The mixture was filtered, and the filtrate concentrated under vacuum. The residue was purified by reversed-phase flash chromatography with the following conditions: column, C18 silica gel; mobile phase, MeCN in Water (lOmmol / L NH4HCO3), 20% to 75% gradient in 30 min; detector, UV 210 nm. This resulted in 4-{2-bromo-8-chloro- [l,2,4]triazolo[l,5-a]pyridin-7-yl}-l-(l-ethoxyethyl)pyrazole (Int-11, 1.8 g, 59%) as a grey solid. LCMS (ESI) calculated for C13H13BrClN5O (M+H)+: 370.64, found: 370.64. RT=0.875 min (analytical method 66).Synthesis of 7-Bromo-8-(2,2,2-trifluoroethoxy)-[l,2,4]triazolo[l,5-a]pyridin-2-amine (Int-12)Step 3

[0244] Step 1 : Into a 500 mL 3-necked round bottle purged and maintained with an inert atmosphere of nitrogen was placed a solution of 2-amino-4-brom opyri din-3 -ol hidrobromide (23.0 g, 85.502 mmol, 1.0 equiv) in DMF (115.0 mL). CS2CO3 (83.6 g, 256.506 mmol, 3.0 equiv) and 2,2,2-trifluoroethyl trifluoromethanesulfonate (21.8 g, 94.052 mmol, 1.1 equiv) were added at room temperature. The mixture was stirred overnight at room temperature. The resulting mixture was diluted with H2O (50.0 ml), and extracted with EA (200.0 ml). The combined organic layers were washed with brine (50.0 ml), dried over anhydrous Na2SO4, fdtered and the fdtrate was concentrated under reduced pressure to afford 4-bromo-3-(2,2,2-trifluoroethoxy)pyridin-2-amine (Int-12a, 22.0 g, crude) as a white solid. The crude was used in the next step without further purification. LCMS (ESI) calculated for C7H6BrF3N2O (M+H)+: 270.96 , found:270.90. RT=1.142 min (analytical method 69).

[0245] Step 2 : Into a 1 L 3-necked round bottle purged and maintained with an inert atmosphere of nitrogen was placed a solution of Int-12a (22.0 g, 81.170 mmol, 1.0 equiv) in ACN (440.0 mL). Ethyl N-carbothioylcarbamate (16.0 g, 121.755 mmol, 1.5 equiv) was added at room temperature and the mixture was stirred overnight at 60 °C. The mixture was then concentrated under reduced pressure to afford ethyl N-({5-chloro-6-[l-(l-ethoxyethyl)-lH-pyrazol-4- yl]pyrimidin-4-yl}carbamothioyl)carbamate (Int-12b, 33.0 g, crude) as a yellow oil. LCMS (ESI) calculated for CnHiiBrFsNsCLS (M+H)+: 401.97, found: 401.65. RT=0.787 min (analytical method 66).

[0246] Step 3 : Into a 2 L 3-necked round bottle purged and maintained with an inert atmosphere of nitrogen was placed a solution of Int-12b (33.0 g, 82.051 mmol, 1.0 equiv) in MeOH (330.0 mL) and EtOH (330.0 mL). Hydroxylamine hydrochloride (17.1 g, 246.153 mmol, 3.0 equiv) and DIEA (31.8 g, 246.153 mmol, 3.0 equiv) were added at room temperature. The mixture was stirred for 1 hour at 60 °C, cooled to room temperature and the volatiles were removed under reduced pressure. The residue was purified by column chromatography (0-40% EA:PE) to afford 7-bromo-8-(2,2,2-trifhjoroethoxy)-[l,2,4]triazolo[l,5-a]pyridin-2-amine (Int-12, 11.0 g, 41.3% yield for 3 steps) as a white solid. LCMS (ESI) calculated for C8H6BrF3N4O (M+H)+: 310.97, found: 311.60. RT=0.694 min (analytical method 66).Synthesis of 7-(l-(l-Ethoxyethyl)-lH-pyrazol-4-yl)-8-(2,2,2-trifluoroethoxy)-[l,2,4]triazolo[l,5- a]pyridin-2-amine (Int-13)

[0247] Into a 500 mL 3-necked round bottle purged and maintained with an inert atmosphere of nitrogen was placed a solution of Int-12 (11.0 g, 35.363 mmol, 1.0 equiv) in dioxane (220.0 mL) and H2O (44.0 mL). l-(l-Ethoxyethyl)-4-(4,4,5,5-tetramethyl-l,3,2-dioxaborolan-2- yl)-lH-pyrazole (10.4 g, 38.899 mmol, 1.1 equiv) and K3PO4 (15.0 g, 70.726 mmol, 2.0 equiv) were added. The reaction mixture was evacuated and flushed 3 times with nitrogen. Pd(dppf)Ch.CH2C12 (1.4 g, 1.768 mmol, 0.05 equiv) was added. The mixture was evacuated and flushed 3 times with nitrogen again and then stirred overnight at 90 °C under nitrogen. The reaction mixture was cooled to room temperature, poured into water (200.0 ml) and extracted with ethyl acetate (500.0 mL). The combined organic layers were washed brine (200.0 ml), dried over Na2SC>4, filtered, and concentrated under reduced pressure. The residue was purified by flash chromatography (0-70% EA:PE) to afford 7-(l-(l-ethoxyethyl)-lH-pyrazol-4-yl)-8-(2,2,2-trifluoroethoxy)-[l,2,4]triazolo[l,5-a]pyridin-2-amine (lnt-13, 9.0 g, 68.7%) as a light-yellow solid. LCMS (ESI) calculated for C15H17F3N6O2 (M+H)+: 371.14 , found: 371.10. RT=0.893 min (analytical method 67).Synthesis of 2-Bromo-7-(l-(l-ethoxyethyl)-lH-pyrazol-4-yl)-8-(2,2,2-trifluoroethoxy)-[l,2,4]triazolo[l,5-a]pyridine (Int-14)

[0248] Step 5 : Into a 500 mL 3-necked round bottle purged and maintained with an inert atmosphere of nitrogen was placed a solution of CuBrc (6.5 g, 29.162 mmol, 1.2 equiv) and tertbutyl nitrite (6.0 g, 58.325 mmol, 2.4 equiv) in ACN (90.0 mL). The mixture was stirred for 5 min at room temperature and treated with a solution of lnt-13 (9.0 g, 24.302 mmol, 1.0 equiv) in ACN(90.0 mL). The mixture was stirred for 30 min at room temperature. The mixture was then filtered and the filtrate was purified by combi-flash with the following conditions [Column: Cl 8; Mobile Phase A: Water (0.1% FA), Mobile Phase B: ACN; Flow rate: 100 mL / min; Gradient: 50% B to 100% B in 30 min; Wave Length: 220 nm; RTl(min): 15.00] to afford 2-bromo-7-(l-(l- ethoxyethyl)-lH-pyrazol-4-yl)-8-(2,2,2-trifluoroethoxy)-[l,2,4]triazolo[l,5-a]pyridine (Int-14, 8.0 g, 75.8% yield) as a white solid. LCMS (ESI) calculated for C15H15BrF3N5O2 (M+H)+: 434.04, found: 434.05. RT=1.201 min (analytical method 65).Synthesis of 7-(l-(l -Ethoxy ethyl )-lH-pyrazol-4-yl)-8-fhioro-[ 1,2, 4]triazolo[l, 5-c]pyrimidin-2- amine (Int-15)

[0249] Step 1 : Into a 500 mL 3-necked round bottom flask purged and maintained with an inert atmosphere of nitrogen was placed a solution of 6-chloro-5-fluoropyrimidin-4-amine (10.0 g, 67.778 mmol, 1.0 equiv), l-(l-ethoxyethyl)-4-(4,4,5,5-tetramethyl-l,3,2-dioxaborolan-2-yl)- IH-pyrazole (19.8 g, 74.556 mmol, 1.1 equiv), and Na2COs (21.6 g, 203.334 mmol, 3.0 equiv) in dioxane (100 mL) and H2O (25 mL). The reaction mixture was degassed 3 times with nitrogen. Then, Pd(dppf)C12 CH2C12 (3.47 g, 4.744 mmol, 0.07 equiv) was added. The mixture was stirred for 2 hours at 90°C under nitrogen. LCMS showed the reaction was completed. The reaction mixture was cooled to room temperature and diluted with ethyl acetate (3 x 40 mL). The combined organic lavers were washed with brine (50 mL), dried over anhydrous Na2SC>4, filtered, and the filtrate was concentrated under reduced pressure. The residue was purified by flash chromatography (Ethyl acetate in petroleum ether 0-60%) to afford 6-( 1-(1 -ethoxy ethyl)- 1H- pyrazol-4-yl)-5-fluoropyrimidin-4-amine (Int-15a, 12.0 g, 66.9%) as a pink solid. LCMS (ESI) calculated for C11H14FN5O (M+H)1: 252.12, found: 252.10; RT=0.643 min (analytical method 65).

[0250] Step 2 : Into a 250 mL 3-necked round bottom flask purged and maintained with an inert atmosphere of nitrogen was placed a solution of Int-15a (6.0 g, 23.879 mmol, 1.0 equiv) in toluene (120 mL). Then, ethyl JV-carbothioylcarbamate (6.26 g, 47.758 mmol, 2.0 equiv) was added at room temperature. The mixture was stirred overnight at 80°C. The resulting mixture was concentrated under reduced pressure to afford ethyl 2V-({6-[l-(l-ethoxyethyl)-lH-pyrazol-4-yl]-5- fluoropyrimidin-4-yl}carbamothioyl)carbamate (Int-15b, 12.7 g, crude) as yellow oil. Thecompound was used in the next step without further purification. LCMS (ESI) calculated for C15H19FN6O3S (M+H)+: 383.12, found: 383.05; RT=0.934 min (analytical method 67).

[0251] Step 3 : Into a 500 mL 3-necked round bottom flask purged and maintained with an inert atmosphere of nitrogen was placed a solution of Int-15b (12.7 g, 23.879 mmol, 1.0 equiv) in MeOH (127 mL) and EtOH (127 mL). Then, NH2OH.HCI (4.9 g, 71.684 mmol, 3.0 equiv) and DIEA (9.2 g, 71.684 mmol, 3.0 equiv) were added at room temperature. The mixture was stirred for 1 hour at 60°C. The reaction mixture was cooled to room temperature. The resulting solution was diluted with EA (150 mL), washed with H2O (3 x 50 mL), and dried with anhydrous Na2SC>4. The resulting mixture was concentrated under vacuum to afford 7-( 1-(1 -ethoxy ethyl)- IH-pyrazol- 4-yl)-8-fhioro-[l,2,4]triazolo[l,5-c]pyrimidin-2-amine (Int-15, 10.0 g, crude) as a yellow solid. The compound was used in the next step without further purification. LCMS (ESI) calculated for C12H14FN7O (M+H)+: 292.12, found: 292.05; RT=0.761 min (analytical method 67).Synthesis of 2-Bromo-7-(l-(l -ethoxy ethyl)- lH-pyrazol-4-yl)-8-fluoro-[ 1,2, 4]triazolo[ 1,5- c]pyrimidine (Int-16)

[0252] Into a 500 mL 3-necked round bottom flask purged and maintained with an inert atmosphere of nitrogen was placed a solution of CuBn (9.2 g, 41.196 mmol, 1.2 equiv) and tertbutyl nitrite (8.5 g, 82.392 mmol, 2.4 equiv) in ACN (100 mL). The mixture was stirred for 5 min at room temperature. Then, Int-15 (10.0 g, 34.330 mmol, 1.0 equiv) in ACN (100 mL) was added. The mixture was stirred for 30 min at room temperature. The mixture was filtered and the filtrate was concentrated and purified by combi-flash [C18 column; Mobile Phase A: Water (0.1% NH4HCO3), Mobile Phase B: ACN; Flow rate: 100 mL / min; Gradient: 40% B to 80% B in 30 min; Wave Length: 210 nm; RT(min): 13.00] to afford 2-bromo-7-(l-(l-ethoxyethyl)-lH-pyrazol-4- yl)-8-fluoro-[l,2,4]triazolo[l,5-c]pyrimidine (Int-16, 1.2 g, 9% yield) as a yellow solid. LCMS(ESI) calculated for Ci2Hi2BrFN6O (M+H)+: 357.02, found: 357.05; RT=0.937 min (analytical method 65).Synthesis of 6-Bromo-5-chloro-[l,2,4]triazolo[l,5-a]pyrazin-2-amine (Int-17)

[0253] Step 1 : Into a 500 ml 3-necks flask purged and maintained with an inert atmosphere of nitrogen was placed a solution of 5-bromo-6-chloropyrazin-2-amine (10 g, 47.975 mmol, 1 equiv) in ACN (200 mL). Ethyl N-carbothioylcarbamate (9.44 g, 71.963 mmol, 1.5 equiv) was added to the mixture and stirred at 60°C for 2 hours. The reaction was cooled to room temperature and the precipitated solids were collected by filtration and washed with ACN (20ml) to afford ethyl N-[(5-bromo-6-chloropyrazin-2-yl) carbamothioyl] carbamate (Int-17a, 10 g, 61.38%). LCMS (ESI) calculated for CsHsBrClN^S (M+H)+: 340.92, found: 341.05. RT= 0.715 min (analytical method 70).[00254J Step 2 : Into a 500 ml 3-necks flask purged and maintained with an inert atmosphere of nitrogen was placed a solution of Int-17a (10 g, 29.447 mmol, 1 equiv) in MeOH (100 mL) and EtOH (100 mL). Hydroxylamine hydrochloride (6.14 g, 88.341 mmol, 3 equiv) and DIEA (11.42 g, 88.341 mmol, 3 equiv) were added and the mixture was stirred at 90°C for 2 hours. The reaction was cooled to room temperature. The precipitated solids were collected by filtration to afford 6- bromo-5-chloro-[l,2,4]triazolo[l,5-a]pyrazin-2-amine (Int-17, 6 g, 82.00%). LCMS (ESI) calculated for CsHsBrClNs (M+H)+: 249.93, found: 249.95. RT= 0.890 min (HALOWA column, analytical method 66).Synthesis of 6-Bromo-5-(2,2,2-trifluoroethoxy)-[l,2,4]triazolo[l,5-a]pyrazin-2-amine (Int-18)120 °C, 2 h lnt-17 Y=45.5% Int-18

[0255] Into a 500 ml 3 -necked round bottle purged and maintained with an inert atmosphere of nitrogen was placed a solution of lnt-17 (14 g, 56.345 mmol, 1 equiv) and 2,2,2- trifluoroethan-l-ol (5.64 g, 56.345 mmol, 1 equiv) in dioxane (140 mL). The mixture was cooled to 0°C and NaH (1.35 g, 56.345 mmol, 1 equiv) was added. The mixture was then stirred at rt for 5 min and stirred at 120°C for 2 hours. The reaction was cooled to room temperature, filtered and the filtrate was concentrated under reduced pressure. The crude was purified by silica gel column chromatography (petroleum ether: ethyl acetate=4: l) to afford 6-bromo-5-(2,2,2-trifluoroethoxy)- [l,2,4]triazolo[l,5-a]pyrazin-2-amine (Int-18, 8 g, 45.50%). LCMS (ESI) calculated for C7H5BrF3N5O (M+H)+: 313.96, found: 313.90. RT= 1.043 min (HALOWA column, analytical method 66).Synthesis of 7-(l-(l-Ethoxyethyl)-lH-pyrazol-4-yl)-8-fluoro-2-iodo-[l,2,4]triazolo[l,5- c]pyrimidine (Int-19)

[0256] Into a 3 L 4-necked round bottle purged and maintained with an inert atmosphere of nitrogen was placed a solution of Int-15 (30.0 g, 102.990 mmol, 1.0 equiv), CH2I2 (55.2 g, 205.980 mmol, 2.0 equiv), and NaNCh (35.5 g, 514.950 mmol, 5.0 equiv) in DCM (600 mL) and H2O (600 mL). The mixture was stirred for 0.5 hours at room temperature. Then, AcOH (123.7 g, 2059.800 mmol, 20.0 equiv) was added and the mixture was stirred for 2 hours at room temperature. The organic phase was then separated and the aqueous layer was extracted with DCM (500 mL x 2). The combined organic layer was concentrated under vacuum and the residue waspurified by silica gel column chromatography, eluted with EA / PE (B:0%~50%, 60min) to 7-(l -(1 - ethoxyethyl)-lH-pyrazol-4-yl)-8-fluoro-2-iodo-[l,2,4]triazolo[l,5-c]pyrimidine (Int-19, 17 g, 41.0%) as an off-white solid. LCMS (ESI) calculated for C12H12FIN6O (M+H)+: 403.01, found: 403.45. RT=1.100 min (analytical method 69).Synthesis of 7-(l -(1 -Ethoxyethyl)- lH-pyrazol-4-yl)-8-methoxy-[ 1,2, 4]triazolo[l,5-c]pyrimidin-

[0257] Step 1 : Into a 500 mL 3-necked round bottle purged and maintained with an inert atmosphere of nitrogen was placed a solution of 6-chloro-5-methoxypyrimidin-4-amine (10.0 g, 62.668 mmol, 1.0 equiv), l-(l-ethoxyethyl)-4-(4,4,5,5-tetramethyl-l,3,2-dioxaborolan-2- yl)pyrazole (16.6 g, 62.668 mmol, 1.0 equiv), and K3PO4 (26.6 g, 125.336 mmol, 2.0 equiv) in dioxane (200 mL) and H2O (40 mL). The reaction mixture was evacuated and flushed 3 times with nitrogen. Then, Pd(dppf)Ch (2.3 g, 3.133 mmol, 0.05 equiv) was added. The mixture was stirred overnight at 90°C under nitrogen. LCMS showed the reaction was completed. The reaction mixture was cooled to room temperature and diluted with ethyl acetate (3 x 100 mL). The combined organic lavers were washed with brine (300 mL), dried over anhydrous NazSCh, filtered, and the filtrate was concentrated under reduced pressure. The residue was purified by flash chromatography (EA: PE=0-60%) to afford 6-(l-(l-ethoxyethyl)-lH-pyrazol-4-yl)-5-methoxypyrimidin-4-amine (Int-20a, 11.0 g, 63.3%) as a yellow solid. LCMS (ESI) calculated for C12H17N5O2 (M+H)+: 264.14, found: 264.20; RT=0.855 min (analytical method 73).[002581 Step 2 : Into a 500 mL 3-necked round bottle purged and maintained with an inert atmosphere of nitrogen was placed a solution of Int-20a (8.0 g, 30.383 mmol, 1.0 equiv) in ACN (160 mL). Then, ethyl A-carbothioylcarbamate (5.9 g, 45.575 mmol, 1.5 equiv) was added at room temperature. The mixture was stirred overnight at 60°C. The resulting mixture was concentrated under reduced pressure to afford ethyl A-({6-[l-(l-ethoxyethyl)-lH-pyrazol-4-yl]-5- methoxypyrimidin-4-yl} carbarn othioyl)carbamate (Int-20b, 15.0 g, crude) as a yellow oil. The crude was used in the next step without further purification. LCMS (ESI) calculated for C16H22N6O4S (M+H)+: 395.14, found: 395.25; RT=0.870 min (analytical method 72).

[0259] Step 3 : Into a 500 mL 3-necked round bottle purged and maintained with an inert atmosphere of nitrogen was placed a solution of Int-20b (15.0 g, 38.028 mmol, 1.0 equiv) in MeOH (150 mL) and EtOH (150 mL). Then, NH2OH.HCI (7.93 g, 114.084 mmol, 3.0 equiv) and DIEA (14.74 g, 114.084 mmol, 3.0 equiv) were added at room temperature. The mixture was stirred for 1 hour at 60°C. The reaction mixture was cooled to room temperature. The resulting solution was diluted with EA (150 mL) and washed with H2O (3 x 50 mL), dried with anhydrous Na2SO4, filtered, and concentrated under vacuum to afford 7-( 1-(1 -ethoxy ethyl)- lH-pyrazol-4-yl)- 8-methoxy-[l,2,4]triazolo[l,5-c]pyrimidin-2-amine (Int-20, 10.0 g, 86.6 %) as a brown oil. The crude was used in the next step without further purification. LCMS (ESI) calculated for C13H17N7O2 (M+H)+: 304.14, found: 304.30; RT=0.743 min (analytical method 72).Synthesis of 2-Bromo-7-( 1 -( 1 -ethoxy ethyl)- lH-pyrazol-4-yl)-8-methoxy-[ 1 ,2,4]triazolo[ 1,5-

[0260] Into a 500 mL 3 -necked round bottle purged and maintained with an inert atmosphere of nitrogen was placed a solution of CuBn (8.8 g, 39.562 mmol, 1.2 equiv) and tertbutyl nitrite (8.2 g, 79.123 mmol, 2.4 equiv) in ACN (100 mb). The mixture was stirred for 5 min at room temperature. Then, Int-20 (10.0 g, 32.968 mmol, 1.0 equiv) in ACN (100 mb) was added. The mixture was stirred for 30 min at room temperature. The mixture was filtered, and the filtrate was purified by combi-flash chromatography [Cl 8 column; Mobile Phase A: Water (0.1% NH4HCO3), Mobile Phase B: ACN; Flow rate: 100 mL / min; Gradient: 40% B to 70% B in 30 min; Wave Length: 210 nm; RT(min): 20.00] to afford 2-bromo-7-(l-(l-ethoxyethyl)-lH-pyrazol-4- yl)-8-methoxy-[l,2,4]triazolo[l,5-c]pyrimidine (Int-21, 3.5 g, 26.0%) as a yellow solid. LCMS (ESI) calculated for CuHuBrNeCh (M+H)+: 369.04, found: 368.95; RT=0.830 min (analytical method 68).Synthesis of 8-Chloro-7-(3-methyl-l-((2-(trimethylsilyl)ethoxy)methyl)-lH-pyrazol-4-yl)-[l,2,4]triazolo[l,5-c]pyrimidin-2-amine (Int-22)

[0261] Step 1 : Into a 250 ml 3-necked round bottle purged and maintained with an inert atmosphere of nitrogen was placed a solution of 3-methyl-4-(4,4,5,5-tetramethyl-l,3,2- dioxaborolan-2-yl)-lH-pyrazole (10.0 g, 48.061 mmol, 1.0 equiv) in DMF (100.0 mb). At room temperature, SEMCI (12.0 g, 72.091 mmol, 1.5 equiv) and CS2CO3 (47.0 g, 144.183 mmol, 3.0 equiv) were added to the mixture and then stirred overnight at room temperature. The resulting mixture was concentrated under reduced pressure to afford 3-methyl-4-(4,4,5,5-tetramethyl-l,3,2- dioxaborolan-2-yl)-l-{[2-(trimethylsilyl)ethoxy]methyl}-lH-pyrazole (Int-22a, 20.0 g, crude) asa brown solid. The product was used in the next step without further purification. LCMS (ESI) calculated for CieHsiBNyOaSi (M+H)+: 339.22, found:339.05, RT=1.311 min (analytical method 74).

[0262] Step 2: Into a 500 ml 3-necked round bottle purged and maintained with an inert atmosphere of nitrogen was placed a solution of Int-22a (20.0 g, 59.114 mmol, 2.5 equiv) in dioxane (200.0 mL) and H2O (50.0 mL). To the mixture were added at room temperature 5,6- dichloropyrimidin-4-amine (3.9 g, 23.646 mmol, 1.0 equiv) and K3PO4 (10.0 g, 47.291 mmol, 2.0 equiv). The mixture was bubbled with N? for 1 min, treated with. Pd(dppf)C12 (0.9 g, 1.182 mmol, 0.05 equiv) and bubbled again with N2 for 1 min. The resulting mixture was stirred overnight at 90 °C and then allowed to cool to room temperature. The resulting mixture was diluted with H2O (10.0 ml), extracted with EA (10.0 ml x 3). The combined organic layers were washed with NaCl (10.0 ml), dried over anhydrous NaySCL, filtered, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography, eluted with EA / PE (1:3) to afford 5- chloro-6-(3-methyl-l-{[2-(trimethylsilyl)ethoxy]methyl}pyrazol-4-yl)pyrimidin-4-amine (Int- 22b, 8.3 g, 51% over 2 steps) as a brown solid. LCMS (ESI) calculated for C14H22ClN5OSi (M+H)+: 340.13, found; 340.05, RT=1.262 min (analytical method 69).

[0263] Step 3: Into a 500 ml 3-necked round bottle purged and maintained with an inert atmosphere of nitrogen was placed a solution of Int-22b (8.3 g, 24.419 mmol, 1.0 equiv) in ACN (166.0 mL) and treated with ethyl N-carbothioylcarbamate (6.4 g, 48.838 mmol, 2.0 equiv) at room temperature. The mixture was stirred for 48 hours at 60 °C and then cooled down to room temperature and concentrated under reduced pressure to afford ethyl N-{[5-chloro-6-(3-methyl-l- {[2-(trimethylsilyl)ethoxy] methyl}pyrazol-4-yl)pyrimidin-4-yl]carbamothioyl}carbamate (Int- 22c, 14.6 g, crude) as a brown solid. The crude product was used in the next step without further purification. LCMS (ESI) calculated for C18H27ClN6O3SSi (M+H)+: 471.13, found: 471.10, RT= 1.295 min (analytical method 74).

[0264] Step 4: Into a 500 ml 3-necked round bottle purged and maintained with an inert atmosphere of nitrogen was placed a solution of Int-22c (14.6 g, 30.995 mmol, 1.0 equiv) in MeOH (146.0 mL) and EtOH (146.0 mL). NH2OH.HCI (6.5 g, 92.985 mmol, 3.0 equiv) and DIEA (12.0 g, 92.985 mmol, 3.0 equiv) were added to the mixture at room temperature and the mixture was stirred for 2 hours at 60 °C. The mixture was cooled to room temperature, the volatiles were removed under reduced pressure and the precipitated solids were collected by filtration and washedwith EtOH (300.0 mL) to afford 8-chloro-7-(3-methyl-l-{[2-(trimethylsilyl)ethoxy]methyl}pyrazol-4-yl)-[l,2,4]triazolo[l,5-c]pyrimidin-2-amine (Int-22, 2.6 g, crude) as a white solid. The crude product was used in the next step without further purification. LCMS (ESI) calculated for C ^ClNvOSi (M+H)+: 380.13, found: 380.15, RT=1.116 min (analytical method 74).Synthesis of 8-Chloro-7-(3-methyl-l-((2-(trimethylsilyl)ethoxy)methyl)-lH-pyrazol-4-yl)-N- (tetrahydro-2H-pyran-4-yl)-[l,2,4]triazolo[l,5-c]pyrimidin-2-amine (Int-23)ep

[0265] Step 5: Into a 20 mL vial was placed at room temperature a solution of Int-22(700.0 mg, 1.842 mmol, 1.0 equiv) in DMF (7.0 mL). The mixture was then cooled to 0 °C with an ice bath and treated with tetrahydro-4H-pyran-4-one (553.4 mg, 5.526 mmol, 3.0 equiv), TFA (1050.4 mg, 9.210 mmol, 5.0 equiv), and STAB (1171.5 mg, 5.526 mmol, 3.0 equiv). The resulting mixture was stirred overnight at room temperature. The mixture was then basified to pH 9 with NaHCCh Solution and extracted with EA (100.0 mL x 2). The combined organic layers were washed with H2O (10.0 mL x 2), dried over anhydrous Na2SO4, fdtered, and concentrated under reduced pressure. The residue was purified by flash chromatography (0-60% EA in PE) to afford 8-chloro-7-(3-methyl-l-{[2-(trimethylsilyl)ethoxy]methyl}pyrazol-4-yl)-N-(oxan-4-yl)- [l,2,4]triazolo[l,5-c]pyrimidin-2-amine (Int-23, 750.0 mg, Y=23.7% over 3 steps) as a white solid. LCMS (ESI) calculated for C2oH3oClN702Si(M+H)+: 464.19, found:446.20, RT=1.219 min (analytical method 74).Synthesis of 7-Chloro-8-cyclobutyl-[l,2,4]triazolo[l,5-c]pyrimidin-2-amine (Int-24)

[0266] Step 1 : Into a 250 mL 3-necked round-bottom flask was placed DCM (100 mL, 10 V) under nitrogen atmosphere. 6-Chloro-5-iodopyrimidin-4-amine (10 g, 39.2 mmol, 1.0 equiv.), TEA (11.88 g, 117.6 mmol, 3.0 equiv.), and di-tert-butyl dicarbonate (10.25 g, 47.04mmol, 1.2 equiv.) were added into the flask at room temperature. The mixture was stirred at room temperature for 6 hours and then filtered. The filtrate was treated with water (30 ml) and extracted with DCM (50 mL x 2). The combined organic layers were washed with water (70 mL x 3), brine (200 mL x 1), dried over with MgSCU, filtered, and concentrated under vacuum to afford crude tert-butyl (6- chloro-5-iodopyrimidin-4-yl)carbamate (Int-24a, 14 g) as a yellow solid. The crude product was used in the next step without further purification. LCMS (ESI) calculated for C9H11CIIN3O2 (M+H)+: 356.56, found: 356.56, RT=1.248 min (analytical method 74).

[0267] Step 2: Into a 100 mL 3-necked round-bottom flask was placed THE (200 ml, 200 vol) under nitrogen atmosphere and then treated at room temperature with Int-24a (1.0 g, 2.81 mmol, 1 equiv ), cyclobutylzinc(II) bromide (17 ml, 4.21 mmol, 1.5 equiv.), and LiCl (118 mg, 2.81 mmol, 1.0 equiv.). The mixture was degassed by a cycle of nitrogen and vacuum for 3 times and treated at room temperature with {l,3-bis[2,6-bis(pentan-3-yl)phenyl]-4,5-dichloro-2,3- dihydro-lH-imidazol-2-yl}dichloro(2 -methyl- llambda4-pyri din- l-yl)palladium (237 mg , 0.28 mmol, 0.1 equiv). The mixture was degassed again by a cycle of nitrogen and vacuum for 3 times and then heated to 70 °C for 16 hours. The mixture was then cooled to room temperature and filtered. The filtrate was treated with water (10 ml) and extracted with EA (10 mL x 2). Thecombined organic layers were washed with water (15 mb x 3), brine (30 mL x 1 ), dried over with Na2SO4, filtered, and concentrated under vacuum. The crude was purified by column chromatography to afford tert-butyl (6-chloro-5-cyclobutylpyrimidin-4-yl)carbamate (Int-24b, 0.7 g, 87% for 2 steps) as a white solid. LCMS (ESI) calculated for C13H18CIN3O2 (M+H)+: 284.76, found: 284.76, RT=1.074 min (analytical method 74).

[0268] Step 3 : Into a 40 mL vial was placed Int-24b (1 g ) under nitrogen atmosphere. Then, HCl / Dioxane (20 ml, 20 vol) was added at room temperature and the resulting mixture was stirred at room temperature until LCMS showed consumption of starting material. The mixture was then filtered and the filtrate was concentrated under vacuum to afford 6-chloro-5- cyclobutylpyrimidin-4-amine (Int-24c, 510 mg, crude ) as a yellow solid. The crude was used in the next step without further purification. LCMS (ESI) calculated for C8H10CIN3 (M+H)+: 184.64, found: 184.64, RT=0.736 min (analytical method 74).

[0269] Step 4 : Into a 50 mL 3-necked round-bottom flask was placed ACN (10 ml, 20 vol) under nitrogen atmosphere. Then, Int-24c (500 mg, 2.73 mmol, 1 equiv.) and O-ethyl carbonisothiocyanatidate (536 mg, 4.09 mmol, 1.5 equiv.) were added at room temperature. The resulting mixture was heated to 90 °C overnight. The mixture was cooled to room temperature and filtered. The filtrate was then diluted with water (30 ml) and extracted with EA (10 mL x 2). The combined organic layers were washed with water (10 mL x 3), brine (20 mL x 1), dried over with Na2SC>4, filtered, and concentrated under vacuum to afford ethyl N-[(6-chloro-5- cyclobutylpyrimidin-4-yl)carbamothioyl]carbamate (Int-24d, 850 mg, 37% over 2 steps). The product was used in the next step without further purification. LCMS (ESI) calculated for Ci2Hi5ClN4O2S(M+H)+: 315.79, found: 315.79, RT=1.011 min (analytical method 62).

[0270] Step 5 : Into a 50 mL 3-necked round-bottom flask were placed MeOH (8.5 ml, 10 vol) and EtOH ( 8.5 ml, 10 vol) under nitrogen atmosphere and treated at room temperature with Int-24d (850 mg, 2.70 mmol, 1 equiv.), HONH3CI (332 mg, 8.12 mmol, 3.0 equiv.), and DIEA (616 mg, 8.12 mmol, 3 equiv.). The resulting mixture was heated to 80 °C and stirred at 80 °C for 2 hours. The mixture was then cooled to room temperature, filtered and the filtrate was diluted with water (5 ml) and extracted with EA (5 mL x 2). The combined organic layers were washed with water (10 mL x 3), brine (20 mL x 1), dried over with Na2SO4, filtered, and concentrated under vacuum. The crude was purified by column chromatography to afford 7-chloro-8- cyclobutyl-[l,2,4]triazolo[l,5-c]pyrimidin-2-amine (Int-24, 230 mg, 38%) as a white solid.LCMS (ESI) calculated for C9H10CIN5 (M+H)+: 224.66, found: 224.66, RT=0.844min (analytical method 28).Synthesis of 8-Cy cl obutyl-7-(l-(l -ethoxy ethyl)-lH-pyrazol-4-yl)-[ 1,2, 4]triazolo[ 1,5- c]pyrimidin-2-amine (Int-25)

[0271] Into a 25 mL 2-necked round-bottom flask were placed dioxane (5 ml, 20 vol) and H2O (0.5 ml, 2 vol) under nitrogen atmosphere and treated at room temperature with lnt-24 (230 mg, 1.03 mmol, 1.0 equiv.), l-(l-ethoxyethyl)-4-(4,4,5,5-tetramethyl-l,3,2-dioxaborolan-2-yl)- IH-pyrazole (410 mg, 1.54 mmol, 1.5 equiv.), and K3PO4 (656 mg, 3.09 mmol, 3 equiv). The mixture was degassed by a cycle of nitrogen and vacuum for 3 times. Pd(dppf)cl2 (75.2 mg, 0.1 mmol, 0.1 equiv) was then added at room temperature and the mixture was degassed again by another cycle of nitrogen and vacuum for 3 times. The resulting mixture was heated to 90 °C and stirred at 90 °C for 12 hours. The mixture was then cooled to room temperature, filtered and the filtrate was diluted with water (5 ml) and extracted with EA (5 mL x 2). The combined organic layers were washed with water (10 mL x 3), brine (20 mL x 1), dried over with Na2SO4, filtered, and concentrated under vacuum. The crude was purified by column chromatography to afford 8- cyclobutyl-7-(l-(l -ethoxyethyl)-lH-pyrazol-4-yl)-[l,2,4]triazolo[l,5-c]pyrimidin-2-amine (Int- 25, 200 mg, 59%). LCMS (ESI) calculated for C16H21N7O (M+H)+: 328.39, found: 328.39, RT=0.815 min (analytical method 28).Synthesis of 8-Chloro-7-(3-methyl-l-((2-(trimethylsilyl)ethoxy)methyl)-lH-pyrazol-4-yl)- [l,2,4]triazolo[l,5-a]pyridin-2-amine (Int-26)

[0272] Step 1 : Into a 250 ml 3-necked round bottom flask purged and maintained with an inert atmosphere of nitrogen was placed a solution of 4-bromo-3-methyl-lH-pyrazole (9.5 g, 59.005 mmol, 1.0 equiv) in THF (475.0 mL) and cooled to 0 °C before being treated with NaH (2.6 g, 64.906 mmol, 1.1 equiv, 60%). The mixture was stirred for 0.5 hours at 0 °C and then treated with SEM-C1 (10.3 g, 61.955 mmol, 1.05 equiv). The mixture was warmed to room temperature and stirred for 2 hours before being quenched with 150.0 ml H2O. The resulting mixture was extracted with EA (150 ml x 3). The combined organic layers were dried over anhydrous Na2SO4, filtered, and concentrated in vacuum. The residue was purified by silica gel column chromatography (EA:PE=0%-10%) to afford 4-bromo-3-methyl-l-{[2-(trimethylsilyl)ethoxy]methyl}-lH-pyrazole (Int-26a, 16.5 g, 96.0% yield) as a yellow oil. LCMS (ESI) calculated for CioHwBrNhOSi (M+H)+: 291.05, found: NA (analytical method 93).[002731 Step 2 : Into a 1000 ml 3 -necked round bottom flask purged and maintained with an inert atmosphere of nitrogen was placed a solution of Int-26a (16.5 g, 56.650 mmol, 1.0 equiv) in THF (320.0 mL). 2-Isopropoxy-4,4,5,5-tetramethyl-l,3,2-dioxaborolane (21.1 g, 113.300 mmol, 2.0 equiv) was added and then the mixture was cooled to -78 °C. n-BuLi (40.8 mL, 101.970 mmol, 1.8 equiv) was added dropwise at -78 °C and the mixture was stirred for 2 hours before being warmed to 0 °C and quenched with 400 ml H2O. The resulting mixture was extracted with EA (400 ml x 3). The combined organic layers were dried over anhydrous NazSCL, filtered, and concentrated in vacuum to afford 3-methyl-4-(4,4,5,5-tetramethyl-l,3,2-dioxaborolan-2-yl)-l-{[2- (trimethylsilyl)ethoxy]methyl}-lH-pyrazole (Int-22a, 28.0 g, crude) as a yellow oil. The crude product was used in the next step without further purification. LCMS (ESI) calculated for Ci6H3iBN2O3Si (M+H)+: 339.22, found: 339.20 (analytical method 65).

[0274] Step 3 : Into a 250 ml 3-necked round bottom flask purged and maintained with an inert atmosphere of nitrogen was placed a solution of Int-22a (13.0 g, 38.424 mmol, 1.0 equiv), 4-bromo-3-chloropyridin-2-amine (3.2 g, 15.370 mmol, 0.4 equiv), and K3PO4 (6.5 g, 30.739 mmol, 0.8 equiv) in dioxane (130.0 mL) and H2O (32.5 mL). Pd(dppf)C12 (562.3 mg, 0.768 mmol, 0.02 equiv) was added to the mixture, warmed to 90 °C and stirred for 2 hours. The reaction was quenched with 150 ml H2O. The resulting mixture was extracted with EA (150 ml x 3). The combined organic layers were dried over anhydrous Na2SO4, filtered, and concentrated in vacuum. The residue was purified by silica gel column chromatography (EA:PE = 0%-40%) to afford 3- chloro-4-(3-methyl-l-{[2-(trimethylsilyl)ethoxy]methyl}-lH-pyrazol-4-yl)pyridin-2-amine (Int- 26b, 5.6 g, 62.8% yield over 2 steps) as a yellow solid. LCMS (ESI) calculated for CisIfeCfNUOSi (M+H)+: 339.13, found: 339.05 (analytical method 66).

[0275] Step 4 : Into a 250 ml 3-necked round bottom flask purged and maintained with an inert atmosphere of nitrogen was placed a solution of Int-26b (5.6 g, 16.475 mmol, 1.0 equiv) and ethyl N-carbothioylcarbamate (3.3 g, 24.713 mmol, 1.5 equiv) in ACN (110.0 mL). The mixture was warmed to 60 °C and stirred for 2 hours. The resulting mixture was concentrated under reduced pressure to afford ethyl N-{[5-chloro-6-(3-methyl-l-{ [2-(trimethylsilyl)ethoxy]methyl}- lH-pyrazol-4-yl)pyrimidin-4-yl]carbamothioyl]carbamate (Int-26c, 9.1 g, crude) as a yellow oil.LCMS (ESI) calculated for CwFEsClNsCLSSi (M+H)+: 470.14, found: 470.00 (analytical method 66).[002761 Step 5 : Into a 250 ml 3-necked round bottom flask purged and maintained with an inert atmosphere of nitrogen was placed a solution of Int-26c (9.1 g, 19.359 mmol, 1.0 equiv), NH2OH.HC1 (4.1 g, 58.077 mmol, 3.0 equiv), andDIEA (7.5 g, 58.077 mmol, 3.0 equiv) in MeOH (91.0 mL) and EtOH (91.0 mL). The mixture was warmed to 60 °C and stirred for 2 hours. The resulting mixture was filtered and the filter cake was washed with MeOH (10.0 mL) to afford 8- chloro-7-(3-methyl-l-{[2-(trimethylsilyl)ethoxy]methyl}-lH-pyrazol-4-yl)-[l,2,4]triazolo[l,5- a]pyridin-2-amine (Int-26, 2.0 g, crude) as an off-white solid. The crude product was used in the next step without further purification. LCMS (ESI) calculated for C 16H23CIN6OSi (M+H)+: 379.14, found: 378.90 (analytical method 66).Synthesis of 8-Chloro-7-(3-methyl-l-((2-(trimethylsilyl)ethoxy)methyl)-lH-pyrazol-4-yl)-N- (tetrahydro-2H-pyran-4-yl)-[l,2,4]triazolo[l,5-a]pyridin-2-amine (Int-27)[00277J Into a 20 mL vial was placed a solution of Int-26 (500 mg, 1.316 mmol, 1.0 equiv) in DMF (5.0 mL). Tetrahydro-4H-pyran-4-one (395.3 mg, 3.948 mmol, 3.0 equiv), TFA (750.3 mg, 6.580 mmol, 5.0 equiv), and NaBH(OAc)s (836.8 mg, 3.948 mmol, 3.0 equiv) were added to the mixture at 0 °C. The resulting mixture was stirred for 2 hours at room temperature and then poured into water (20.0 ml). The resulting mixture was extracted with EA (20 mL x 3). The combined organic layers were washed with brine (30.0 ml), dried over Na2SO4, filtered, and concentrated under reduced pressure to afford 8-chloro-7-(3-methyl-l-((2- (trimethylsilyl)ethoxy)methyl)-lH-pyrazol-4-yl)-N-(tetrahydro-2H-pyran-4-yl)-[l,2,4]triazolo[l,5-a]pyridin-2-amine (Int-27, 680 mg, crude) as a white solid. The crude material was used in the next step without further purification. LCMS (ESI) calculated for C21H31ClN6H32O2Si (M+H)+: 463.20, found: 463.30 (analytical method 73).Synthesi s of 2-Bromo-7-(3 , 5-dimethyl- 1 -(tetrahy dro-2H-pyran-2-yl)- 1 H-py razol-4-y 1)-[ 1 , 2, 4]tri azolof 1 ,5-a]pyridine (Int-28)

[0278] Step 1 : Into a 100 mL 3 -necked round bottle purged and maintained with an inert atmosphere of nitrogen was placed a solution of CuBn (1.3 g, 5.633 mmol, 1.2 equiv) and tertbutyl nitrite (1.2 g, 11.266 mmol, 2.4 equiv) in ACN (20 mL). To the mixture was added at room temperature 7-bromo-[l,2,4]triazolo[l,5-a]pyridin-2-amine (1.0 g, 4.694 mmol, 1.0 equiv) and the resulting mixture was stirred for 0.5 hours at room temperature. The resulting mixture was filtered; the filter cake was washed with ACN (10 mL) and the filtrate was concentrated under reduced pressure. The residue was purified by combi-flash (Column: C18; Mobile Phase A: Water (0.1% FA), Mobile Phase B: ACN; Flow rate: 100 mL / min; Gradient: 15% B to 55% B in 30 min; Wave Length: 210 / 254 nm; RT(min): 25) to afford 2,7-dibromo-[l,2,4]triazolo[l,5-a]pyridine (Int-28a, 320 mg, 22% yield) as a yellow solid. LCMS (ESI) calculated for C6H3Br2N3 [M+H]1; 277.92, found: 277.90, RT = 0.875 min (analytical method 74).

[0279] Step 2 : Into an 8 mL vail was placed a solution of Int-28a (150.0 mg, 0.542 mmol, 1.0 equiv) in dioxane (3 mL) and H2O (0.6 mL). 3,5-Dimethyl-l-(oxan-2-yl)-4-(4,4,5,5- tetramethyl-l,3,2-dioxaborolan-2-yl)-lH-pyrazole (165.9 mg, 0.542 mmol, 1.0 equiv) and K3PO4 (229.96 mg, 1.084 mmol, 2.0 equiv) were added at room temperature. The mixture was bubbled with N2 for 2 minutes and treated at room temperature with Pd(dppf)C12 (19.9 mg, 0.027 mmol, 0.05 equiv). The mixture was then stirred for 2 hours at 90 °C. The residue was purified by reverse phase flash chromatography (C18 column; Mobile Phase A: Water (0.1% NH4HCO3), Mobile Phase B: ACN; Flow rate: 100 mL / min; Gradient: 35% B to 55% B in 30 min; Wave Length: 210 / 254 nm; RT(min): 10) to afford 4-{2-bromo-[l,2,4] triazolo[l,5-a]pyridin-7-yl}-3,5- dimethyl-l-(oxan-2-yl)-lH-pyrazole (Int-28, 120 mg, 58% yield) as a yellow oil. LCMS (ESI)calculated for CieHisBrNsO [M+H]+; 376.07, found: 376.05, RT = 1.003 min (analytical method 65).Synthesis of 7-Bromo-N-(tetrahydro-2H-pyran-4-yl)-[l,2,4]triazolo[l,5-a]pyridin-2-amine (Int-Y=77% lnt-29

[0280] Into a 250 ml 3-necks flask bottle purged and maintained with an inert atmosphere of nitrogen was placed a solution of 7-bromo-[l,2,4]triazolo[l,5-a]pyridin-2-amine (6.0 g, 28.16 mmol, 1.0 equiv) in DMF (60.0 mL) and cooled to 0 °C. Tetrahydro-4H-pyran-4-one (8.5 g, 84.49 mmol, 3.0 equiv), TFA (32.4 g, 281.64 mmol, 10.0 equiv), and STAB (18.0 g, 84.49 mmol, 3.0 equiv) were added and the mixture was stirred for 3 hours at room temperature. The residue was purified by reverse phase flash chromatography (C18 column, 0.1% NH4HCO3 / ACN, 18% ACN to 55% ACN, RT=14.5 min) to afford 7-bromo-N-(oxan-4-yl)- [1,2,4] triazolo[l,5-a]pyridin-2- amine (lnt-29, 6.5 g, 77% yield) as a white solid. LCMS (ESI) calculated for CnHi3BrN4O (M+H)+: 297.03, found: 296.95; RT = 0.811 min (analytical method 65).Synthesis of N-(Tetrahydro-2H-pyran-4-yl)-7-(4,4,5,5-tetramethyl-l,3,2-dioxaborolan-2-yl)- [l,2,4]triazolo[l,5-a]pyridin-2-amine (Int-30).

[0281] Into a 40 mL vial were added at room temperature Int-29 (1 g, 3.365 mmol, 1 equiv), 4,4,5,5-tetramethyl-2-(4,4,5,5-tetramethyl-l,3,2-dioxaborolan-2-yl)-l,3,2-dioxaborolane (1.71 g, 6.730 mmol, 2 equiv), cataCXium-A-Pd-G3 (0.37 g, 0.505 mmol, 0.15 equiv), bis(adamantan-l-yl)(butyl)phosphane (0.18 g, 0.505 mmol, 0.15 equiv), KOAc (0.50 g, 5.048 mmol, 1.5 equiv), and dioxane (16 mL). The resulting mixture was stirred for 3 hours at 90 °C under nitrogen atmosphere. The mixture was cooled down to room temperature and purified by reverse phase flash chromatography (Cl 8 column; mobile phase, MeCN in Water (10 mmol / L NH4HCO3), 10% to 30% gradient in 10 min; detector, UV 210 nm) to afford A-(oxan-4-yl)-7- (4,4,5,5-tetramethyl-l,3,2-dioxaborolan-2-yl)-[l,2,4]triazolo[l,5-a]pyridin-2-amine (Int-30, 840 mg, 72.51%) as a white solid. LCMS (ESI) calculated for C17H25BN4O3 [M+H]+; 345.2, found: 345.25, RT = 0.761 min (analytical method 73).Synthesis of 5-Isopropyl-4-(4,4,5,5-tetramethyl-l,3,2-dioxaborolan-2-yl)-l-((2-(trimethylsilyl)ethoxy)methyl)-lH-pyrazole (lnt-31)step 1 step 2 lnt-31a lnt-31

[0282] Step 1. Into a 100 mL 3 -necks flask bottle purged and maintained with an inert atmosphere of nitrogen was placed a solution of 4-bromo-3-isopropyl-2H-pyrazole (2.5 g, 13.22 mmol, 1.0 equiv) in DMF (20.0 mL), followed by [2-(chloromethoxy)ethyl]trimethylsilane (3.3 g, 19.83 mmol, 1.5 equiv) and CS2CO3 (12.9 g, 39.67 mmol, 3.0 equiv). The mixture was stirred overnight at room temperature and then extracted with EA (50.0 mL x 3). The combined organic layers were washed with H2O (50.0 mL x 3), dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure. The crude was purified by column chromatography (PE / EA 0—1.6%) to afford 4-bromo-5-isopropyl-l-{[2-(trimethylsilyl)ethoxy]methyl}-lH-pyrazole (Int- 31a, 3 g, 75% yield) as a colorless oil. LCMS (ESI) calculated for Ci2H23BrN2OSi (M+H)+: 319.08, found: 319.05; RT=1.357 min (analytical method 62).

[0283] Step 2. Into a 20 mL vial purged and maintained with an inert atmosphere of nitrogen was placed a solution of Int-31a (500.0 mg, 1.56 mmol, 1.0 equiv) in THF (10.0 mL). 2-isopropoxy-4,4,5,5-tetramethyl-l,3,2-dioxaborolane (582.68 mg, 3.13 mmol, 2.0 equiv) was added and the mixture was cooled at -70 °C before being treated dropwise with n-BuLi (1.2 mb). The mixture was then stirred for 2 hours at -70 °C, quenched at the same temperature with saturated aqueous NH4CI solution. The resulting mixture was extracted with EA (3 x 15.0 ml), dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure to afford 5-isopropyl-4- (4,4,5,5-tetramethyl-l,3,2-dioxaborolan-2-yl)-l-{ [2-(trimethylsilyl)ethoxy]methyl }-lH-pyrazole (Int-31, 580.0 mg, crude) as a yellow oil. The crude product was used in the next step directly without further purification. LCMS (ESI) calculated for CisHisB^OiSi (M+H)+: 367.25, found: 367.25; RT=1.503 min (analytical method 62).Synthesis of 8-Chloro-7-(l-(l-ethoxyethyl)-lH-pyrazol-4-yl)-N-((3S,4S)-3-fluorotetrahydro-2H- pyran-4-yl)-[ 1 , 2, 4]tri azolof 1 ,5-c]pyrimidin-2-amine (Int-32)

[0284] Into a 2 L 3-necked round-bottom flask was placed a solution of Int-9 (60.0 g,161.4 mmol, 1.0 equiv.), (3S,4S)-3-fluorotetrahydro-2H-pyran-4-amine hydrochloride (28.8 g,242.5 mmol, 1.5 equiv.), and CS2CO3 (157.6 g, 485.1mmol, 3 equiv.) in dioxane (1.2 L, 20 V) under nitrogen atmosphere. The mixture was degassed by a cycle of nitrogen and vacuum for 3 times. Then, Pd2(dba)3 (14.8 g, 16.2 mmol, 0.1 equiv.) and xantphos (18.7 g, 32.3 mmol, 0.2 equiv.) were added. The mixture was degassed again by a cycle of nitrogen and vacuum for 3 times and the resulting mixture was heated to 90°C and stirred for 16 hours upon which LCMS showed complete conversion. The mixture was cooled to room temperature, diluted with water (1 L) and extracted with EA (1.5 L x 2). The combined organic layers were washed with brine (1 L), dried over with anhydrous Na2SO4, fdtered, and concentrated under vacuum. The crude was purified by column chromatography to afford 8-chloro-7-(l-(l-ethoxyethyl)-lH-pyrazol-4-yl)-N-((3S,4S)-3- fluorotetrahydro-2H-pyran-4-yl)-[l,2,4]triazolo[l,5-c]pyrimidin-2-amine (Int-32, 50 g, 37% over 2 steps) as a yellow solid. LCMS (ESI) calculated for C17H21CIFN7O2 (M+H)+: 410.14, found: 410.20, RT=0.904 min. (Analytical method 65).Synthesis of 4-Iodo-5-isobutyl-l-(4-methoxybenzyl)-lH-pyrazole (Int-33)PMB-CI (1.2 eq) PMB DMF (10 V)Cs2CO3(2 eq)N-vK60 °C, 1 h1Y = 64.71%Int-33

[0285] Into an 8 mL vial were added at room temperature 4-iodo-5-isobutyl-lH-pyrazole (100 mg, 0.400 mmol, 1 equiv), CS2CO3 (260.57 mg, 0.800 mmol, 2 equiv), PMB-CI (326.28 mg, 0.4800 mmol, 1.2 equiv), and DMF (1 mL). The resulting mixture was stirred for 1 hour at 60 °C under nitrogen atmosphere. The residue was purified by reverse phase flash chromatography (Cl 8 column; mobile phase, MeCN in Water (lOmmol / L NH4HCO3), 30% to 50% gradient in 20 min; detector, UV 210 nm) to afford 4-iodo-5-isobutyl-l-(4-methoxybenzyl)-lH-pyrazole (Int-33, 95 mg, 64.17%) as a brown solid. LCMS (ESI) calculated for C15H19IN2O [M+H]+; 371.05, found: 371.10, RT = 1.371 min (analytical method 73).Synthesis of 5-Chloro-6-(l-(l-ethoxyethyl)-lH-pyrazol-4-yl)-N-((3S,4S)-3-fluorotetrahydro-2H- pyran-4-yl)-[l,2,4]triazolo[l,5-a]pyrazin-2-amine (Int-34)

[0286] Into a 8 ml vial purged and maintained with an inert atmosphere of nitrogen was placed a solution of Int-2 (300 mg, 0.807 mmol, 1.0 equiv) and (3S,4S)-3-fluorotetrahydro-2H- pyran-4-amine hydrochloride (96.18 mg, 0.807 mmol, 1.0 equiv) in dioxane (6 mL). Then, at room temperature, CS2CO3 (789.1 mg, 2.421 mmol, 3.0 equiv), XantPhos (186.8 mg, 0.323 mmol, 0.4 equiv), and Pd2(dba)3CHCh (167.1 mg, 0.161 mmol, 0.2 equiv) were added. The mixture was bubbled with N2 for 1 min and stirred overnight at 100 °C. The reaction was cooled to room temperature and purified by reversed-phase flash chromatography (C18 column, A:0.1%NH4HCO3 in water, B: ACN; 20% B-50% B, 30 min) to afford 5-chloro-6-(l-(l-ethoxyethyl)-lH- pyrazol-4-yl)-N-((3S,4S)-3-fluorotetrahydro-2H-pyran-4-yl)-[l,2,4]triazolo[l,5-a]pyrazin-2- amine (Int-34 ,40 mg, 12.1%) as a yellow solid. LCMS (ESI) calculated for C17H21CIFN7O2 (M+H)+: 410.14, found: 410.20, RT=1.039 min. (analytical method 62).Synthesis of 6-Bromo-N-(tetrahydro-2H-pyran-4-yl)-5-(2,2,2-trifluoroethoxy)-[ 1 ,2,4]triazolo[ 1 ,5-a]pyrazin-2-amine (Int-35)Y=91% lnt-18 lnt-35

[0287] Into a 20 mL vial purged and maintained with an inert atmosphere of nitrogen was placed a solution of lnt-18 (500.0 mg, 1.602 mmol, 1.0 equiv) and tetrahydro-4H-pyran-4-one (802.1 mg, 8.010 mmol, 5.0 equiv) in DMF (9 mL). The mixture was then cooled to 0°C and treated with TFA (1.83 g, 16.020 mmol, 10.0 equiv) followed by STAB (1.02 g, 4.806 mmol, 3.0 equiv). The mixture was stirred at room temperature for 4 hours, cooled to 0°C, and neutralized to pH 7 with aqueous NaOH solution. The reaction mixture was extracted with EA (50 mL x 3), washed with brine, dried with anhydrous Na2SC>4, filtered, and concentrated to afford 6-bromo-N- (tetrahydro-2H-pyran-4-yl)-5-(2,2,2-trifluoroethoxy)-[l,2,4]triazolo[l,5-a]pyrazin-2-amine (Int- 35, 680 mg, 91%) as a yellow solid. LCMS (ESI) calculated for Ci2Hi3BrF3N5O2 (M+H)+: 396.02, found: 395.90, RT=0.748 min. (analytical method 68).Example 3 - Synthesis of Compounds 1-173Synthesis of N-((3S,4S)-3-Fluorotetrahydro-2H-pyran-4-yl)-7-(lH-pyrazol-4-yl)-8-(2,2,2- trifluoroethoxy)-[ 1 ,2,4]triazolo[ 1 ,5-c]pyrimidin-2-amine (Compound 2)Compound 2a Compound 2

[0288] Step 1 : Into a 500 mL 3-necked round-bottom flask purged and maintained with an inert atmosphere of nitrogen was placed a solution of Int-7 (8.0 g, 18.433 mmol, 1.0 equiv) in dioxane (160 mL). Then, (3S,4S)-3-fluorotetrahydro-2H-pyran-4-amine hydrochloride (8.6 g, 55.3 mmol, 3.0 equiv) and CS2CO3 (18.0 g, 55.3 mmol, 3.0 equiv) were added to the mixture at room temperature. The mixture was bubbled with N2 for 1 min, treated at room temperature with { 1,3- bis[2,6-bis(pentan-3-yl)phenyl]-4,5-dichloro-2,3-dihydro-lH-imidazol-2-yl}dichloro(2-methyl- llambda4-pyridin-l-yl)palladium (4.6 g, 5.53 mmol, 0.3 equiv), and then bubbled again with N2 for 1 min. The resulting mixture was stirred for 4 hours at 100 °C, cooled down to room temperature and filtered. The filtrate was purified by combi-flash chromatography (Cl 8 column. ACNikbO with 0.1% NH4HCO3 20%~80%, 30 min) to afford 7-(l-(l-ethoxyethyl)-lH-pyrazol- 4-yl)-N-((3S,4S)-3-fluorotetrahydro-2H-pyran-4-yl)-8-(2,2,2-trifluoroethoxy)- [l,2,4]triazolo[l,5-c]pyrimidin-2-amine (Compound 2a, 4.5 g, 51.7% yield) as a yellow solid. LCMS (ESI) calculated for C19H23F4N7O3 [M+H]1; 474.18, found: 474.10, RT = 0.969 min (analytical method 67).

[0289] Step 2 : Into a 250 mL 3-necked round-bottom flask purged and maintained with an inert atmosphere of nitrogen was placed a solution of Compound 2a (4.5 g, 9.514 mmol, 1.0 equiv) in DCM (90 mL). Then, TFA (13.5 mL) was added at room temperature and the resulting mixture was stirred at room temperature for 2 hours. The mixture was concentrated and the residue was purified by combi-flash chromatography (C18 column, ACFFFLO with 0.1% NH4HCO3 20%~50%, 30 min) to afford N-((3S,4S)-3-fluorotetrahydro-2H-pyran-4-yl)-7-(lH-pyrazol-4-yl)- 8-(2,2,2-trifluoroethoxy)-[l,2,4]triazolo[l,5-c]pyrimidin-2-amine (Compound 2, 1.5 g, 39.3% yield) as a white solid. LCMS (ESI) calculated for C15H15F4N7O2 [M+H]+; 402.12, found: 402.10, RT = 1.513 min (analytical method 25); 'H-NMR (400 MHz, DMSO-c / <, ppm}. 5 13.24 (s, 1H), 9.20 (s, 1H), 8.26 (s, 1H), 8.11 (s, 1H), 7.36 (d, J = 7.6 Hz, 1H), 5.41 - 5.29 (m, 2H), 4.86 (d,, / =49.3 Hz, 1H), 4.09 - 3.80 (m, 3H), 3.67 - 3.44 (m, 2H), 2.00 -1.85 (m, 1H), 1.78 - 1.68 (m, 1H);19F-NMR (376 MHz, DMSO4 p»i): 8 -73.01, -203.90.Synthesis of 8-Cyclobutyl-7-(lH-pyrazol-4-yl)-N-(tetrahydro-2H-pyran-4-yl)-[l,2,4]triazolo[l,5- c]pyrimidin-2-amine (Compound 3)

[0290] Step 1 : Into a 10 mL 2-necked round-bottom flask were placed DMF (4 ml, 20 vol) and TFA (185 mg, 1.83 mmol, 3.0 equiv) under nitrogen atmosphere. Int-25 (200 mg, 0.61mmol, 1 equiv.), NaBH(OAc)3 (259 mg, 1.22 mmol, 2.0 equiv.), and tetrahydro-4H-pyran-4-one (123 mg, 1.22 mmol, 2 equiv.) were added at room temperature and the mixture was stirred at room temperature for 17 hours. The mixture was then filtered, the filtrate diluted with water (5 ml) and extracted with EA (5 mL x 2). The combined organic layers were washed with water (10 mL x 3), brine (20 mL x 1), dried over with Na2SC>4, filtered, and concentrated under vacuum to afford 8- cyclobutyl-7-(l -(1 -ethoxy ethyl)-lH-pyrazol-4-yl)-N-(tetrahydro-2H-pyran-4-yl)-[l,2,4]triazolo[l,5-c]pyrimidin-2-amine (Compound 3a, 191 mg, crude). The product was used in the next step without further purification. LCMS (ESI) calculated for C21H29N7O2 (M+H)+: 413.51, found: 413.51, RT=1.125 min (analytical method 93).

[0291] Step 2 : Into an 8 mL vial was placed DCM (3.8 ml) under nitrogen atmosphere. Compound 3a (190 mg) was added into the flask at room temperature, followed by TFA (0.57 ml). The resulting mixture was stirred at room temperature until LCMS showed complete reaction. The mixture was then filtered and the filtrate concentrated under vacuum. The crude was purified by Flash-Prep-HPLC (Xselect CSH Prep Fluoro-phenyl Column, 30*150 nm, 5 pm; Mobile Phase A: Water (0.1% FA), Mobile Phase B: ACN; Flow rate: 60 mL / min; Gradient: 5% B to 5% B in 2 min, 5% B to 15% B in 2.5 min, 15% B to45% B in 10 min; Wave Length: 220 / 254 nm; RT(min): 8.62, Detector, UV 210 nm) to afford 8-cyclobutyl-7-(lH-pyrazol-4-yl)-N-(tetrahydro-2H-pyran- 4-yl)-[l,2,4]triazolo[l,5-c]pyrimidin-2-amine (Compound 3, 66.3 mg, 32% over 2step) as an off- white solid. LCMS (ESI) calculated for C17H21N7O (M+H)+: 340.20, found: 340.20, RT = 1.448 min (analytical method 27); 1H NMR (400 MHz, DMSO-tL, ppm) 8 9.21 (s, 1H), 7.98 (s, 2H),7.10 (s, 1H), 4.01 (p, J = 9.1 Hz, 1H), 3.89 (dt, J= 11.5, 3.5 Hz, 2H), 3.71 (tt, J = 10.2, 4.0 Hz, 1H), 3.40 (td, J= 11.6, 2.2 Hz, 2H), 2.95 (pd, J= 9.3, 2.6 Hz, 2H), 2.26 - 2.14 (m, 2H), 1.95 (dddd, 7= 21.1, 13.5, 10.6, 5.3 Hz, 4H), 1.62 - 1.47 (m, 2H).Synthesis of N-((3S,4S)-3-Fluorotetrahydro-2H-pyran-4-yl)-6-(lH-pyrazol-4-yl)-5-(2,2,2- trifluoroethoxy )-[ 1 ,2,4]triazolo[ 1 , 5-a]pyrazin-2-amine (Compound 12)

[0292] Step 1 : Into a 8 ml vial was placed a solution of lnt-34 (100.0 mg, 0.244 mmol, 1.0 equiv) and trifluoroethanol (29.3 mg, 0.293 mmol, 1.2 equiv) in dioxane (1 mL). The reaction was cooled to 0°C andNaH (9.8 mg, 0.244 mmol, 1.0 equiv, 60%wt) was added. The mixture was stirred for 5 min at room temperature and then stirred at 90°C for 2 hours. The reaction was cooled to room temperature, quenched by the addition of H2O (5 ml) and extracted with EA (50 mL x 3). The combined organic layers were washed with brine, dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure. The crude was purified by reversed-phase flash chromatography (C18 column; mobile phase, ACN in Water (lOmmol / L NH4HCO3), 40% to 70% gradient in 30 min; detector, UV 210 nm) to afford 6-(l-(l-ethoxyethyl)-lH-pyrazol-4-yl)-N- ((3S,4S)-3-fluorotetrahydro-2H-pyran-4-yl)-5-(2,2,2-trifluoroethoxy)-[l,2,4]triazolo[l,5- a]pyrazin-2-amine (Compound 12a, 80.0 mg, 69.3%) as a yellow oil. LCMS (ESI) calculated for C19H23F4N7O3 (M+H)+: 474.18, found: 474.30, RT=1.101 min. (analytical method 62).

[0293] Step 2 : In to a 8 ml vial was added a solution of Compound 12a (130.0 mg, 0.275 mmol, 1.0 equiv) and TFA (0.39 mL) in DCM (2.6 mL), the mixture was stirred for 0.5 hours at room temperature. The resulting mixture was concentrated under reduced pressure and the residue was purified by reversed-phase flash chromatography (XSelect CSH Fluoro Phenyl column, 30*150 mm, 5pm; Mobile Phase A: Water(10mmol / L NH4HCO3), Mobile Phase B: ACN; Flow rate: 60 mL / min mL / min; Gradient: 38% B to 52% B in 10 min; Wave Length: 254nm / 220nm nm; RT(min): 8.18) to afford N-((3S,4S)-3-fluorotetrahydro-2H-pyran-4-yl)-6-(lH-pyrazol-4-yl)-5- (2,2,2-trifluoroethoxy)-[l,2,4]triazolo[l,5-a]pyrazin-2-amine (Compound 12, 75.9 mg, 68.2%) as a white solid. LCMS (ESI) calculated for C15H15F4N7O2 (M+H)+: 402.12, found: 402.05,RT=1.444 min. (analytical method 12); 'H-NMR (400 MHz, DMSO-rL, ppm . 8 13.12 (bs, 1H), 8.77 (s, 1H), 8.11 (bs, 1H), 8.04 (bs, 1H), 7.41 (d, J= 7.6 Hz, 1H), 5.30 (q, J= 9.0 Hz, 2H), 4.86 (d, J = 49.2 Hz, 1H), 4.12 - 3.86 (m, 3H), 3.79 - 3.45 (m, 2H), 1.94 (qd, J = 12.4, 4.6 Hz, 1H), 1.73 (d, J= 12.5 Hz, 1H);19F-NMR (376 MHz, DMS0 / H: 8 -72.59, -203.99.Synthesis of 8-(2-Azabicyclo[2.1.1]hexan-2-yl)-N-((3S,4S)-3-fluorotetrahydro-2H-pyran-4-yl)-7-(lH-pyrazol-4-yl)-[l,2,4]triazolo[l,5-c]pyrimidin-2-amine (Compound 19)

[0294] Step 1 : Into a 100 mL 3-necked round-bottom flask were added at room temperature Int-32 (2.5 g, 6.100 mmol, 1.0 equiv), 2-azabicyclo[2.1.1]hexane hydrochloride (1094 mg, 9.150 mmol, 1.0 equiv), { l,3-bis[2,6-bis(pentan-3-yl)phenyl]-4,5-dichloro-2,3- dihydro- lH-imidazol-2-yl } dichloro(2 -methyl- 1 lambda4-pyridin- 1 -yl)palladium (513.08 mg, 0.610 mmol, 0.1 equiv), CS2CO3 (5962.28 mg, 18.300 mmol, 3.0 equiv), and dioxane (25 mL). The resulting mixture was stirred overnight at 90 °C under nitrogen atmosphere. The residue was purified by reverse phase flash chromatography (Cl 8 column; mobile phase, MeCN in Water (10 mmol / L NH4HCO3), 30% to 50% gradient in 20 min; detector, UV 210 nm.) to afford 8-(2- azabicyclo[2. l.l]hexan-2-yl)-7-(l-(l-ethoxy ethyl)- lH-pyrazol-4-yl)-N-((3S,4S)-3- fluorotetrahydro-2H-pyran-4-yl)-[l,2,4]triazolo[l,5-c]pyrimidin-2-amine (Compound 19a, 1.4 g, 50.3%) as a brown solid. LCMS (ESI) calculated for C22H29FN8O2 (M+H)+: 457.24, found: 457.30, RT = 1.153 min (analytical method 73).

[0295] Step 2: Into a 20 mL vial charged with Compound 19a (400 mg, 0.876 mmol, 1.0 equiv) was added at room temperature DCM (8 mL) and TFA (2 mL) and the mixture was put under nitrogen atmosphere. The resulting mixture was then stirred for 2 hours at room temperature. The residue was purified by reverse phase flash chromatography (Cl 8 column; mobile phase, MeCN in Water (10 mmol / L NH4HCO3), 25% to 40% gradient in 20 min; detector, UV 210 nm) to afford 8-(2-azabicyclo[2.1. l]hexan-2-yl)-N-((3S,4S)-3-fluorotetrahydro-2H-pyran-4-yl)-7- (lH-pyrazol-4-yl)-[l,2,4]triazolo[l,5-c]pyrimidin-2-amine (Compound 19, 253.4 mg, 74.5%) as a white solid. LCMS (ESI) calculated for C18H2iFN8O (M+H)+: 385.19, found: 385.20, RT = 1.110min (analytical method 10); 'H NMR (400 MHz, DMSO-d6. ppm): 8 13.02 (s, 1H), 9.00 (s, 1H), 8.12 (d, 7= 27.6 Hz, 2H), 7.07 (d, 7= 7.6 Hz, 1H), 4.85 (d, 7= 49.1 Hz, 1H), 4.53 (dt, 7= 6.6, 1.7 Hz, 1H), 4.00 (t, 7= 12.4 Hz, 1H), 3.93 - 3.77 (m, 2H), 3.65 - 3.42 (m, 2H), 2.87 - 2.79 (m, 1H), 2.00 - 1.82 (m, 3H), 1.76 - 1.64 (m, 1H), 1.57 (dd, 7 = 4.5, 1.8 Hz, 2H);19F NMR (376 MHz, DMSO4,p / >ffl): 8 -203.98.Synthesis of 8-(3,3-Difluoropyrrolidin-l-yl)-N-((3S,4S)-3-fluorotetrahydro-2H-pyran-4-yl)-7-( lH-pyrazol-4-yl)-[ 1 ,2,4]triazolo[ 1 ,5-c]pyrimidin-2-amine (Compound 65)

[0296] Step 1: Into a 20 mL vial were added Int-32 (200 mg, 0.488 mmol, 1.0 equiv), 3,3- difluoropyrrolidine hydrochloride (105 mg, 0.732 mmol, 1.5 equiv), { l,3-bis[2,6-bis(pentan-3- yl)phenyl]-4,5-dichl oro-2, 3-dihy dro-lH-imidazol -2-yl }dichloro(2-methyl-llambda4-pyridin-l - yl)palladium (41.05 mg, 0.049 mmol, 0.1 equiv), CS2CO3 (476.98 mg, 1.464 mmol, 3.0 equiv), and dioxane (4 mL) at room temperature. The resulting mixture was stirred overnight at 90 °C under nitrogen atmosphere. The mixture was allowed to cool down to room temperature and the crude product was purified by reverse phase flash chromatography (Cl 8 column; mobile phase, MeCN in Water (10 mmol / L NH4HCO3), 30% to 50% gradient in 20 min; detector, UV 210 nm) to afford 8-(3 , 3 -difluoropyrrolidin- 1 -yl)-7-( 1 -( 1 -ethoxy ethyl)- 1 H-pyrazol-4-yl)-N-((3 S,4 S)-3 - fluorotetrahydro-2H-pyran-4-yl)-[l,2,4]triazolo[l,5-c]pyrimidin-2-amine (Compound 65a, 120 mg, 51.2%) as a white solid. LCMS (ESI) calculated for C21H27F3N8O2 [M+H]+; 481.22, found: 481.05, RT = 0.806 min (analytical method 66).

[0297] Step 2 : Into an 8 mL vial were added Compound 65a (100 mg, 0.208 mmol, 1.0 equiv), TFA (0.5 mL) and DCM (2 mL) at room temperature and placed under nitrogen atmosphere. The resulting mixture was stirred for 2 hours and the residue was purified by reverse phase flash chromatography (XSelect Prep OBD C18 Column, 30*150 mm, 5pm; Mobile Phase A: Water (10 mmol / L NH4HCO3), Mobile Phase B: ACN; Flow rate: 60 mL / min; Gradient: 5% B to 5% B in 2 min, 5% B to 18% B in 2.5 min, 18% B to 48% B in 10 min; Wave Length: 220 / 254nm; RT(min): 8.58) to afford 8-(3,3-difluoropyrrolidin-l-yl)-N-((3S,4S)-3-fluorotetrahydro-2H- pyran-4-yl)-7-(lH-pyrazol-4-yl)-[l,2,4]triazolo[l,5-c]pyrimidin-2-amine (Compound 65, 49.6 mg, 57.7%) as a white solid. LCMS (ESI) calculated for C17H19F3N8O [M+H]~; 409.17, found: 409.20, RT = 1.495 min (analytical method 18); 'H NMR (400 MHz, DMSO-tfc, j>pw): 8 13.18 (s, 1H), 9.23 (s, 1H), 8.38 (s, 1H), 8.19 (s, 1H), 7.34 (d, J= 7.6 Hz, 1H), 4.86 (d, J= 49.2 Hz, 1H), 4.05 - 3.76 (m, 5H), 3.64 - 3.47 (m, 4H), 2.62 - 2.50 (m, 2H), 1.94 (qd, J= 12.5, 4.5 Hz, 1H), 1.77 - 1.67 (m, 1H);19F NMR (376 MHz, DMSO-^ / ipm): 8 -93.71, -203.99.Synthesis of 8-(3,3-Difluoropyrrolidin-l-yl)-7-(5-methyl-lH-pyrazol-4-yl)-N-(tetrahydro-2H- pyran-4-yl)-[l,2,4]triazolo[l,5-c]pyrimidin-2-amine (Compound 72)

[0298] Step 1: Into a 20 mL vial were placed a solution of Int-23 (300.0 mg, 0.646 mmol, 1.0 equiv) and 3, 3 -difluoropyrrolidine (83.0 mg, 0.775 mmol, 1.2 eq) in dioxane (6.0 mL). CS2CO3 (842.5 mg, 2.584 mmol, 4.0 equiv) was then added at room temperature. The mixture was bubbled with N2 for 1 min, treated with PPd-PEPPSI-IPentCl 2-methylpyridine (o-picoline) (163.1 mg, 0.194 mmol, 0.3 equiv) and bubbled again with N2 for 1 min. The resulting mixture was stirred overnight at 90 °C. The mixture was then allowed to cool to room temperature, diluted with H2O (10 ml), and extracted with EA (10 ml). The combined organic layers were washed with NaCl (10 ml), dried over anhydrous Na2SCU, fdtered, and concentrated under reduced pressure to afford 8- (3,3-difluoropyrrolidin-l-yl)-7-(3-methyl-l-((2-(trimethylsilyl)ethoxy)methyl)-lH-pyrazol-4-yl)- N-(tetrahydro-2H-pyran-4-yl)-[l,2,4]triazolo[l,5-c]pyrimidin-2-amine (Compound 72a, 150 mg, crude) as a brown solid. The crude product was used in the next step without further purification. LCMS (ESI) calculated for C24H36F2N8O2Si (M+H)+: 535.27, found: 535.30, RT=1.492 min (analytical method 69).

[0299] Step 2: Into an 8 ml vial was placed at room temperature a solution of Compound 72a (1.0 equiv) in TBAF (1 M in THF, 3.0 mL). The resulting mixture was stirred for 2 hours at 70 °C and then cooled down to room temperature. After filtration, the filtrate was concentratedunder reduced pressure and was purified by combi-flash (Xselect CSH Prep Cis OBD Colum, 19*250 nm, 5 pm; Mobile Phase A: Water (10 mmol / L NH4HCO3), Mobile Phase B: ACN; Flow rate: 25 mL / min; Gradient: isocratic 5% B in 2 min, 22% B to 52% B in 10 min; Wave Length: 254 nm / 220 nm; RT(min): 8.8) to afford 8-(3,3-difluoropyrrolidin-l-yl)-7-(5-methyl-lH-pyrazol- 4-yl)-N-(tetrahydro-2H-pyran-4-yl)-[l,2,4]triazolo[l,5-c]pyrimidin-2-amine (Compound 72, 6.5 mg, 7.3%) as a white solid. LCMS (ESI) calculated for C18H22F2N8O [M+H]+; 405.19, found: 405.20, RT = 1.114 min (analytical method 10);1H-NMR (400 MHz, DMSO-J4, ppm}. 8 12.77 (bs, 1H), 9.15 (s, 1H), 7.96 (s, 1H), 7.07 (d, J= 7.7 Hz, 1H), 3.94 - 3.84 (m, 2H), 3.84 - 3.75 (m, 2H), 3.74 - 3.63 (m, 1H), 3.50 - 3.40 (m, 4H), 2.49 - 2.34 (m, 5H), 1.97 - 1.86 (m, 2H), 1.59 - 1.46 (m, 2H);19F-NMR (376 MHz, DMSO-de, ppm}. 8 -95.18Synthesi s of 7-(3 , 5 -Dimethyl- 1 H-pyrazol -4-yl)-N-((3 S,4 S)-3 -fluorotetrahy dro-2H-pyran-4-yl)- [l,2,4]triazolo[l,5-a]pyridin-2-amine (Compound 76)

[0300] Step 1 : Into an 8 mL vail was placed a solution of Int-28 (90.0 mg, 0.239 mmol, 1.0 equiv) and (3S,4S)-3-fluorotetrahydro-2H-pyran-4-amine hydrochloride (28.5 mg, 0.239 mmol, 1.0 equiv) in dioxane (1.8 mL). CS2CO3 (54.6 mg, 0.717 mmol, 3.0 equiv) was added to the mixture at room temperature. The mixture was bubbled with N2 for 2 minutes, then treated with Pd-PEPPSI-IPentCl 2-methylpyridine (o-picoline) (20.2 mg, 0.024 mmol, 0.1 equiv) and the mixture was stirred for 2 hours at 100 °C. The resulting solution was diluted with EA (10 ml), washed with H2O (3 x 3 ml), dried with anhydrous Na2SO4, filtered, and concentrated under vacuum to afford 7-(3,5-dimethyl-l-(tetrahydro-2H-pyran-2-yl)-lH-pyrazol-4-yl)-N-((3S,4S)-3- fluorotetrahydro-2H-pyran-4-yl)-[l,2,4]triazolo[l,5-a]pyridin-2-amine (Compound 76a, 250 mg, crude) as a brown solid. The crude product was used in the next step without further purification. LCMS (ESI) calculated for C21H27FN6O2 [M+H]+; 415.22, found: 415.20, RT = 0.676 min (analytical method 65).

[0301] Step 2: Into a 2 mL vail was placed Compound 76a (250.0 mg, 0.603 mmol, 1.0 equiv) in 1,4-di oxane (3.75 mL). HC1 (gas) in 1,4-di oxane (3.75 mL) was then added to the mixture at room temperature and the resulting mixture was stirred for 2 hours. The residue was purified by HPLC (YMC-Actus Triart C18 ExRS 30*150 mm column, 5 pm; Mobile Phase A: Water (10 mmol / L NH4HC03+0.05%NH3.H20), Mobile Phase B: ACN; Flow rate: 60 mL / min; Gradient: 35% B to 60% B in 8 min; Wave Length: 254 nm; RT(min): 7) to afford 7-(3,5-dimethyl-lH- pyrazol-4-yl)-N-((3S,4S)-3-fluorotetrahydro-2H-pyran-4-yl)-[l,2,4]triazolo[l,5-a]pyridin-2- amine (Compound 76, 27 mg, 13% yield) as a white solid. LCMS (ESI) calculated for Ci6Hi9FN6O [M+H]+; 331.17, found: 387.15, RT = 1.282 min (analytical method 86); 'H-NMR (400 MHz, DMSO-d6,ppm: 8 12.48 (s, 1H), 8.57 (d, J= 6.9 Hz, 1H), 7.25 (d, J= 1.8 Hz, 1H), 6.86 (dd, J = 7.0, 1.9 Hz, 1H), 6.70 (d, J= 8.1 Hz, 1H), 4.83 (d, J= 49.4 Hz, 1H), 4.07 - 3.83 (m, 3H), 3.67 - 3.42 (m, 2H), 2.26 (s, 6H), 1.90 (dd, J= 12.6, 4.5 Hz, 1H), 1.71 (dd, J= 13.6, 4.6 Hz, 1H);19F-NMR (376 MHz, DMSO-dd6,ppm): 8 -204.24.Synthesis of 6-(3-Fluoro-lH-pyrazol-4-yl)-N-(tetrahydro-2H-pyran-4-yl)-5-(2,2,2- trifluoroethoxy)-[l,2,4]triazolo[l,5-a]pyrazin-2-amine (Compound 80)80°C, o / n Y=8% lnt-35 Compound 80

[0302] Into a 8 ml vial purged and maintained with an inert atmosphere of nitrogen was placed a solution of Int-35 (150.0 mg, 0.379 mmol, 1.0 equiv) and 3-fluoro-4-(4,4,5,5-tetramethyl- l,3,2-dioxaborolan-2-yl)-lH-pyrazole (96.3 mg, 0.455 mmol, 1.2 equiv) in dioxane (3 mL) and H2O (0.3 mL). Then, K3PO4 (120.6 mg, 0.569 mmol, 1.5 equiv) was added to the mixture followed by the addition of Pd(dppf)C12CH2C12 (30.8 mg, 0.038 mmol, 0.1 equiv). The mixture was bubbled with N2 for 1 min and then stirred at 80°C overnight. The reaction was cooled to room temperature, filtered and the crude product was purified by reverse phase flash chromatography (Xbridge Prepphenyl OBD Colum, 30*150nm, 5pm; Mobile Phase A: Water (10 mmol / L NH4HCO3), Mobile Phase B: MEOH; Flow rate: 80 mL / min mL / min; Gradient: 15% B to 50% B in 30 min; Wave Length: 254nm / 220nm nm; RT(min): 18) to afford 6-(3 -fluoro- lH-pyrazol-4-yl)-N-(tetrahydro- 2H-pyran-4-yl)-5-(2,2,2-trifluoroethoxy)-[l,2,4]triazolo[l,5-a]pyrazin-2-amine (Compound 80, 13.4 mg, 8.7%) as a white solid. LCMS (ESI) calculated for C15H15F4N7O2 (M+H)+: 402.12, found: 402.15, RT=1.072 min. (analytical method 10); 'H-NMR (400 MHz, DMSO-d6,ppm}. 8 8.77 (s, 1H), 7.97 (s, 1H), 7.29 (d, J = 7.6 Hz, 1H), 5.19 (q, J = 8.9 Hz, 2H), 3.89 (dt, J = 11.5, 3.6 Hz, 2H), 3.85 - 3.73 (m, 1H), 3.46 - 3.36 (m, 3H), 1.97 - 1.88 (m, 2H), 1.60 - 1.46 (m, 2H);19F-NMR (376 MHz, DMSO-t / 6,j>pm): 8 -72.94, -131.98.Synthesis of 8-(2-Azabicyclo[2.1.1]hexan-2-yl)-7-(3-methyl-lH-pyrazol-4-yl)-N-(tetrahydro-2H- pyran-4-yl)-[l,2,4]triazolo[l,5-a]pyridin-2-amine (Compound 86)

[0303] Step 1 : Into a 20 mL vial was placed a solution of Int-23 (650.0 mg, 1.401 mmol,1.0 equiv) in dioxane (13.0 mL) and treated at room temperature with 2-azabicyclo[2.1.1]hexane hydrochloride (151.4 mg, 1.821 mmol, 1.3 equiv) and CS2CO3 (1825.6 mg, 5.604 mmol, 4.0 equiv). The mixture was bubbled with N2 for 1 min. Pd-PEPPSI-IPentCl 2-methylpyridine (o- picoline) (353.5 mg, 0.420 mmol, 0.3 equiv) was added and the mixture was bubbled again with N2 for 1 min. The resulting mixture was stirred overnight at 90 °C. The resulting mixture was then purified by Combi-Flash chromatography (C18 column; Mobile Phase A: Water (10 mmol / L NH4HCO3), Mobile Phase B: ACN; Flow rate: 80 mL / min; Gradient: 30% B to 70% B in 20 min Wave Length: 254 nm / 210 nm; RT(min): 17) to afford 8-(2-azabicyclo[2.1.1]hexan-2-yl)-7-(3- methyl-l-((2-(trimethylsilyl)ethoxy)methyl)-lH-pyrazol-4-yl)-N-(tetrahydro-2H-pyran-4-yl)- [l,2,4]triazolo[l,5-a]pyridin-2-amine (Compound 86a, 300.0 mg, 14.9% yield over 4 steps) as a white solid. LCMS (ESI) calculated for C26H39N7O2Si (M+H) : 510.29, found: 510.30 (analytical method 73).

[0304] Step 2 : Into a 20 ml vial was placed a solution of Compound 86a (300.0 mg, 0.589 mmol, 1.0 equiv) in DCM (3.0 mL) and cooled to 0 °C before being treated with TFA (3.0 mL). The mixture was then warmed to room temperature and stirred for 2 hours. The resulting mixture was purified by Combi-Flash chromatography (Cl 8 column; Mobile Phase A: Water (10 mmol / L NH4HCO3), Mobile Phase B: ACN; Flow rate: 80 mL / min; Gradient: 20% B to 60% B in 30 min Wave Length: 254 nm / 210 nm; RT(min): 17) to afford 8-(2-azabicyclo[2.1.1]hexan-2-yl)-7-(3- methyl-lH-pyrazol-4-yl)-N-(tetrahydro-2H-pyran-4-yl)-[l,2,4]triazolo[l,5-a]pyridin-2-amine (Compound 86, 85 mg, 37.5% yield) as a white solid. LCMS (ESI) calculated for C20H25N7O [M+H]+; 380.21, found: 380.25, RT = 1.119 min (analytical method 10); 'H-NMR (400 MHz, DMSO-tZ<j,ppw): 8 12.60 (bs, 1H), 8.01 (d, J= 6.7 Hz, 1H), 7.57 (s, 1H), 6.57 (d, J= 6.6 Hz, 1H), 6.34 (d, J= 7.8 Hz, 1H), 5.34 (dt, J= 6.7, 1.8 Hz, 1H), 3.93 - 3.82 (m, 2H), 3.69 - 3.54 (m, 1H), 3.34-3.32 (m, 2H), 2.72 - 2.61 (m, 3H), 2.14 (s, 3H), 1.97 - 1.87 (m, 2H), 1.87 - 1.75 (m, 2H), 1.58 - 1.43 (m, 2H), 1.31 - 1.20 (m, 2H).Synthesis of 7-(3-Methyl-lH-pyrazol-4-yl)-N-(tetrahydro-2H-pyran-4-yl)-[l,2,4]triazolo[l,5- a]pyridin-2-amine (Compound 87)

[0305] Step 1 : Into a 20 ml vial was placed a solution of Int-29 (250.0 mg, 0.84 mmol, 1.0 equiv) in dioxane (5.0 mL) and H2O (1.0 mL). The mixture was then treated at room temperature with tert-butyl 3-methyl-4-(4,4,5,5-tetramethyl-l,3,2-dioxaborolan-2-yl)pyrazole-l- carboxylate (259.2 mg, 0.84 mmol, 1.0 equiv) and K3PO4 (357.1 mg, 1.68 mmol, 2.0 equiv). The mixture was then bubbled with N2 for 2 min, treated with Pd(dppf)C12.CH2C12 (34.2 mg, 0.04 mmol, 0.05 equiv) and bubbled again with N2 for 2 min before being stirred for 2 hours at 90 °C. The resulting solution was extracted with water and EA (10.0 mL x 3). The organic layers were combined, washed with brine, dried with Na2SO4, filtered, and concentrated under reduced pressure to afford tert-butyl 3 -methyl -4-(2-((tetrahy dro-2H-pyran-4-yl )amino)- [ 1 ,2,4]triazol o[ 1,5-a]pyridin-7-yl)-lH-pyrazole-l -carboxylate (Compound 87a, 400 mg, crude) as a yellow oil. The crude product was used in the next step without further purification. LCMS (ESI) calculated for C20H26N6O3 (M+H)+:399.21, found:399.20; RT = 0.941 min (analytical method 65).

[0306] Step 2 : Into a 20 mL vial was placed a solution of Compound 87a (400.0 mg, 1.0 mmol, 1.0 equiv) in dioxane (4.0 mL). HCl(gas) in 1,4-dioxane (2.0 mL) was then added at room temperature and the mixture was stirred for 2 hours before being concentrated under vacuum. The residue was purified by reverse phase flash chromatography (YMC-Actus Triart C 18 ExRS30* 150 mm column, 5 pm; Mobile Phase A: Water (10 mmol / L NH4HC03+0.05%NH3.H20), Mobile Phase B: ACN; Flow rate: 60 mL / min; Gradient: 2% B to 30% B in 12 min; Wave Length: 254nm / 220nm; RT(min): 9.35) to afford 7-(3-methyl-lH-pyrazol-4-yl)-N-(tetrahydro-2H-pyran- 4-yl)-[l,2,4]triazolo[l,5-a]pyridin-2-amine (Compound 87, 100 mg, 30% yield over two steps) as a white solid. LCMS (ESI) calculated for CisHisNeO [M+H]+; 299.15, found: 299.10, RT = 1.068 min; (analytical method 85); 1H-NMR (400 MHz, DMSO-d6,ppm): 5 8.53 (d, 7.2 Hz, 1H), 8.00 (s, 1H), 7.35 (s, 1H), 6.95 (dd, J=7.2 Hz, 1.6 Hz, 1H), 6.50 (d, J=8.0 Hz, 1H), 3.88 - 3.78 (m, 2H), 3.70 - 3.60 (m, 1H), 2.45 (s, 3H), 1.91 - 1.83 (m, 2H), 1.64 - 1.38 (m, 2H).Synthesis of 7-(5-Isopropyl-lH-pyrazol-4-yl)-N-(tetrahydro-2H-pyran-4-yl)-[l,2,4]triazolo[l,5- a]pyridin-2-amine (Compound 88)

[0307] Step 1 : Into a 40 ml vial was placed a solution of Int-29 (350 mg, 1.17 mmol, 1.0 equiv) in dioxane (7.0 mL) and H2O (1.4 mL). The mixture was then treated at room temperature with Int-31 (647.3 mg, 1.76 mmol, 1.5 equiv) and K3PO4 (505.5 mg, 2.35 mmol, 2.0 equiv). The mixture was bubbled with N2 for 2 min, treated with Pd(dppf)C12.CH2Ch (48.5 mg, 0.06 mmol, 0.05 equiv) and bubbled again with N2 for 2 min before being stirred for 2 hours at 90 °C. The resulting solution was extracted with water and EA (10.0 ml x 3). The organic layers were combined, washed with brine, dried with Na2SO4, filtered, and concentrated under reducedpressure to afford 7-(5-isopropyl-l-((2-(trimethylsilyl)ethoxy)methyl)-lH-pyrazol-4-yl)-N- (tetrahydro-2H-pyran-4-yl)-[l,2,4]triazolo[l,5-a]pyridin-2-amine (Compound 88a, 500 mg, crude) as a yellow oil. The crude product was used in the next step without further purification. LCMS (ESI) calculated for C23H36N6O2Si (M+H)+: 457.27, found: 457.45; RT=1.009 min (analytical method 74).

[0308] Step 2 : Into a 20 mL vial was placed a solution of Compound 88a (400 mg, 0.87 mmol, 1.0 equiv) in DCM (4.0 mL) and treated at room temperature with TFA (2.0 mL). The mixture was then stirred for 1 hour at room temperature. The resulting mixture was concentrated under reduced pressure and the residue purified by reverse phase flash chromatography (XSelect Prep OBD C18 Column, 30*150 mm, 5 pm; Mobile Phase A: Water (0.1% FA), Mobile Phase B: ACN; Gradient: 5% B to 5% B in 2 min, 5% B to 9% B in 2.5 min, 9% B to 25% B in 10 min; Wave Length: 220 / 254 nm; RT(min): 10.75) to afford 7-(5-isopropyl-lH-pyrazol-4-yl)-N- (tetrahydro-2H-pyran-4-yl)-[l,2,4]triazolo[l,5-a]pyridin-2-amine (Compound 88, 87 mg, 30% yield over two steps) as a white solid. LCMS (ESI) calculated for C17H22N6O [M+H]1; 327.19, found: 327.20, RT = 1.261 min (analytical method 12); 'H-NMR (400 MHz, DMSO-d6,ppm). 5 12.94 (s, 1H), 8.56 (d, .7=7.2 Hz, 1H), 7.88 (s, 1H), 7.30 (d, J=0.8 Hz, 1H), 6.95 (dd, 7=7.6 Hz, 1.6 Hz, 1H), 6.57 (d, .7=7.6 Hz, 1H), 3.89 - 3.80 (m, 2H), 3.72 - 3.68 (m, 1H), 3.45 - 3.21 (m, 3H), 1.99 - 1.84 (m, 2H), 1.62 - 1.43 (m, 2H), 1.20 (d, .7=2,4 Hz, 6H).Synthesis of 7-(5-Isobutyl-lH-pyrazol-4-yl)-N-(tetrahydro-2H-pyran-4-yl)-[l,2,4]triazolo[l,5- a]pyridin-2-amine (Compound 89)

[0309] Step 1: Into an 8 mL vial were added Int-30 (54 mg, 0.157 mmol, 1 equiv.), Int- 33 (70 mg, 0.189 mmol, 1.2 equiv), K3PO4 (66.04 mg, 0.289 mmol, 1.84 equiv), Pd(dppf)C12CH2Cl2 (38.50 mg, 0.047 mmol, 0.05 equiv), dioxane (1.4 mL) and H2O (0.28 mL) at room temperature. The resulting mixture was degassed and fdled with nitrogen and then stirredovernight at 90 °C. The residue was purified by reverse phase flash chromatography (Cl 8 column; mobile phase, MeCN in Water (10 mmol / L NH4HCO3), 20% to 50% gradient in 20 min; detector, UV 254 nm) to afford 7-(5-isobutyl-l-(4-methoxybenzyl)-lH-pyrazol-4-yl)-N-(tetrahydro-2H- pyran-4-yl)-[l,2,4]triazolo[l,5-a]pyridin-2-amine (Compound 89a, 17 mg, 23.43%) as a white solid. LCMS (ESI) calculated for C26H32N6O2 [M+Hf; 461.26, found: 461.35, RT = 1.152 min (analytical method 73).

[0310] Step 2: Into an 8 mL vial were added at room temperature Compound 89a (270 mg, 0.586 mmol, 1 equiv) and trifluoroacetaldehyde (5 mL, 0.010 mmol, 0.28 equiv). The resulting mixture was stirred for 2 hours at 80 °C under nitrogen atmosphere. The residue was purified by reverse phase flash chromatography (Cl 8 column; mobile phase, MeCN in Water (10 mmol / L NH4HCO3), 20% to 50% gradient in 20 min; detector, UV 210 nm.) to afford 7-(5-isobutyl-lH- pyrazol-4-yl)-N-(tetrahydro-2H-pyran-4-yl)-[l,2,4]triazolo[l,5-a]pyridin-2-amine (Compound 89, 94.3 mg, 46.64%) as a white solid. LCMS (ESI) calculated for CisEbiNeO [M+H]+; 341.20, found: 341.25, RT = 1.495 min (analytical method 10); 'H-NMR (400 MHz, DMSO-t / e, ppm). 3 8.54 (d, .7= 7.0 Hz, 1H), 7.92 (s, 1H), 7.34 (d, J= 1.8 Hz, 1H), 6.97 (dd, J= 7.0, 1.9 Hz, 1H), 6.55 (d, J= 8.0 Hz, 1H), 6.05 (s, 4H), 3.88 (d, J= 11.8 Hz, 2H), 3.69 (d, J= 6.8 Hz, 2H), 3.43 - 3.34 (m, 4H), 2.72 (d, J = 7.2 Hz, 3H), 2.00 - 1.84 (m, 3H), 1.56 - 1.46 (m, 2H), 0.86 (d, J = 6.6 Hz, 6H).

[0311] A list of compounds synthesized as described above is shown in Table 1 below.Table 1. Exemplary Compounds

[0312] The compounds from Table 2 below can be synthesized from the appropriate intermediates and building blocks using one or more reaction conditions from Scheme 1 or Scheme 2 as described above.Table 2: Additional Examples of Compounds of Formula (I)

[0313] The compounds from Table 3 below can be synthesized from the appropriate intermediates and building blocks using one or more reaction conditions from Scheme 3 as described above.Table 3. Examples of Compounds From Formula (II)Example 4 - NanoBRET Target Engagement Assay

[0314] HEK293T cells were transfected with the CDK NanoLuciferase fusions and respective cyclin binding partner or empty plasmid (Promega, Madison, WI). The cells were grown overnight at 37°C, 5%CO2, 100% humidity and plated at a density of 2 x 105 cells / mL in assay media in a 384-well white, tissue culture treated plate. Following overnight incubation, the following probes were added to the cells: K-10 was added for CDK1, 2, 5 and 7, K-7 for CDK6 and K8 for CDK9 at the determined EC50. Following probe addition, compounds were added to the cells. After 2 hours of incubation at 37°C, 5%, CO2, 100% humidity, detection reagents were added according to manufacturer’s instructions (Promega, Madison, USA). The donor (450 nM) and the acceptor (610 nM) emission wavelengths were read on an EnVision plate reader (PerkinElmer, Waltham, MA). The ratio of the raw count of the acceptor was divided by the raw count for the donor and then normalized by the high and low control values. The Ki was calculated using the Cheng-Prusoff equation (Biochem. Pharmacol, 1973, 22(23):3099).

[0315] Biological activity data are reported in Table 4 as follows:

[0316] A Ki below 20 nM

[0317] B Ki between 20 nM and below 250 nM

[0318] C Ki between 250 nM and below 1000 nM

[0319] D Ki between 1000 nM and below 5000 nM

[0320] E Ki equal or greater than 5000 nMTable 4. Biological Activity of Selected Compounds

[0321] As various changes can be made in the above-described subject matter without departing from the scope and spirit of the present disclosure, it is intended that all subject matter contained in the above description, or defined in the appended claims, be interpreted as descriptive and illustrative of the present disclosure. Many modifications and variations of the present disclosure are possible in light of the above teachings. Accordingly, the present description is intended to embrace all such alternatives, modifications, and variances which fall within the scope of the appended claims.

[0322] All patents, applications, publications, test methods, literature, and other materials cited herein are hereby incorporated by reference in their entirety as if physically present in this specification.

Claims

WHAT IS CLAIMED IS:

1. A compound having a structure selected from the group consisting of:Ĭ0.a pharmaceutically acceptable salt thereof.

2. A compound having a structure as shown in Tables 1-3, or a pharmaceutically acceptable salt thereof.

3. A pharmaceutical composition comprising the compound of claim 1 or claim 2, or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier.

4. A pharmaceutical dosage form comprising the compound of claim 1 or claim 2, or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition of claim 3.

5. A method of targeting cyclin-dependent kinase 2 (CDK2) in a cell, the method comprising contacting the cell with an effective amount of the compound of claim 1 or claim 2, or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition of claim 3, or the pharmaceutical dosage form of claim 4.

6. The method of claim 5, wherein the cell is a mammalian cell.

7. The method of claim 5 or claim 6, wherein the cell is a tumor cell.

8. The method of claim 7, wherein the tumor cell is a metastatic tumor cell.

9. A method of inhibiting CDK2 comprising contacting the CDK2 with an effective amount of the compound of claim 1 or claim 2, or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition of claim 3, or the pharmaceutical dosage form of claim 4.

10. The method of any one of claims 5-9, wherein the contacting is in a subject.

11. A method of treating a subject having, or at risk of developing, a disease or disorder associated with CDK2, comprising administering to the subject a therapeutically effective amount of the compound of claim 1 or claim 2, or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition of claim 3, or the pharmaceutical dosage form of claim 4.

12. A method of treating a CDK2-mediated disorder or disease in a subject, the method comprising administering a therapeutically effective amount of the compound of claim 1 or claim 2, or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition of claim 3, or the pharmaceutical dosage form of claim 4.

13. The method of claim 11 or claim 12, wherein the disorder is cancer.

14. The method of claim 13, wherein the cancer is selected from the group consisting of adrenal gland cancer, anal cancer, appendiceal cancer, ovarian cancer, uterine cancer, gastric cancer, breast cancer, lung cancer, bladder cancer, cervical cancer, stomach cancer, sarcoma cancer, liver cancer, esophageal cancer, laryngeal cancer, multiple myeloma, colorectal cancer, rectal cancer, skin cancer, pancreatic cancer, brain or spinal cord cancer, leukemia or lymphoma.

15. The method of any one of claims 5-14, wherein the compound is at least two-fold more selective for CDK2 over CDK1, CDK 5, CDK 6, CDK7, and / or CDK9.

16. A method of treating a cancer in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of the compound of claim 1 or claim 2, or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition of claim 3, or the pharmaceutical dosage form of claim 4.

17. The method of claim 16, wherein the cancer is characterized by a genomic alteration.

18. The method of claim 16, wherein the cancer is characterized by overexpression of cyclin El and / or cyclin E2.

19. The method of claim 17, wherein the genomic alteration is characterized by one or more of amplification of CCNE1 and / or CCNE2, RBI loss of function mutation, FBXW7 loss of function mutation, MYC amplification, or KRAS mutation.

20. The method of claim 18, wherein the overexpression of cyclin El and / or cyclin E2 is the overexpression of RNA and / or protein encoded by CCNE1 and / or CCNE2.

21. The method of claim 18, wherein greater than two copies of CCNE1 and / or CCNE2 are present.

22. The method of claim 19, wherein the RBI loss of function mutation comprises functiondisabling point mutations in the RBI gene, RBI gene deletion, and / or epigenetic repression of RBI transcription.

23. The method of claim 19, wherein the MYC is selected from c-MYC, L-MYC, and N- MYC.

24. The method of any one of claims 13-23, wherein the cancer is metastatic cancer.

25. The method of claim 24, wherein the cancer is characterized by metastasis to the brain.

26. The method of claim 25, wherein the cancer selected from the group consisting of adrenal gland cancer, anal cancer, appendiceal cancer, ovarian cancer, uterine cancer, gastric cancer, breast cancer, lung cancer, bladder cancer, cervical cancer, stomach cancer, sarcoma cancer, liver cancer, esophageal cancer, laryngeal cancer, multiple myeloma, colorectal cancer, rectal cancer, skin cancer, pancreatic cancer, brain or spinal cord cancer, leukemia or lymphoma.

27. The method of claim 14 or claim 26, wherein the adrenal gland cancer is adrenocortical carcinoma or pheochromocytoma.

28. The method of claim 14 or claim 26, wherein the breast cancer is HR-positive / HER2- negative breast cancer; HR-positive / HER2-positive breast cancer; triple negative breast cancer (TNBC); or inflammatory breast cancer.

29. The method of claim 14 or claim 26, wherein the breast cancer is drug-naive breast cancer, endocrine resistant breast cancer, trastuzumab resistant breast cancer, breast cancer demonstrating primary or acquired resistance to CDK4 / CDK6 inhibition, platinum-resistant breast cancer or platinum -refractory breast cancer.

30. The method of claim 14 or claim 26, wherein the breast cancer is advanced or metastatic breast cancer.

31. The method of claim 14 or claim 26, wherein the liver cancer is hepatocellular carcinoma.

32. The method of claim 14 or claim 26, wherein the lung cancer is small cell lung cancer, non-small cell lung cancer, or large cell lung cancer.

33. The method of claim 32, wherein the non-small cell lung cancer is squamous cell carcinoma or adenocarcinoma.

34. The method of claim 14 or claim 26, wherein the laryngeal cancer is laryngeal squamous cell carcinoma.

35. The method of claim 14 or claim 26, wherein the skin cancer is melanoma, basal cell carcinoma, squamous cell carcinoma, or Merkel cell cancer.

36. The method of claim 14 or claim 26, wherein the uterine cancer is endometrial cancer or uterine sarcoma.

37. The method of claim 36, wherein the endometrial cancer is platinum-resistant or platinum-refractory endometrial cancer.

38. The method of claim 14 or claim 26, wherein the ovarian cancer is platinum-resistant or platinum-refractory ovarian cancer.

39. The method of claim 14 or claim 26, wherein the brain cancer is selected from glioma, astrocytoma, meningioma, glioblastoma multiforme, medulloblastoma, ependymoma, oligodendroglioma, craniopharyngioma, pituitary adenoma, Schwannoma, anaplastic astrocytoma, germ cell tumor, primitive neuroectodermal tumor, chordoma, haemangioblastoma, optic nerve glioma, subependymoma, and germinoma.

40. The method of any one of claims 10-39, wherein the subject is a human.

41. The method of claim 40, wherein the subject is an adult.

42. The method of claim 40, wherein the subject is pediatric subject 0 to 18 years of age.

43. The method of any one of claims 10-42, wherein the compound, the pharmaceutical composition, or the pharmaceutical dosage form is administered orally, intravenously, or subcutaneously.

44. The method of any one of claims 5-43, wherein the method further comprises administering an effective amount of a second therapeutic agent.

45. The method of claim 44, wherein the second therapeutic agent is an anti -neoplastic agent.

46. The method of claim 45, wherein the anti-neoplastic agent is selected from the group consisting of an antibody, an antibody-drug conjugate, a DNA synthesis inhibitor, a platinumbased agent, topoisomerase I inhibitor, topoisomerase II inhibitor, an alkylating agent, an antimicrotubule agent, an anti-mitotic agent, a taxane-related anti-neoplastic agent, an antimetabolite, an anti-tumor plant alkaloid, a DNA-intercalating agent, an anti-estrogen agent, a protein kinase inhibitor, a phosphatidylinositol 3 -kinase inhibitor, an immunomodulator, a histone deacetylase inhibitor, a KRAS inhibitor, an immunotherapeutic agent, or an epigenetic modulator.

47. The method of claim 46, wherein the antibody-drug conjugate comprises sacituzumab govitecan, mirvetuximab soravtansine, trastuzumab deruxtecan, enfortumab vedotin, or tisotumab vedotin.

48. The method of claim 46, wherein the DNA synthesis inhibitor comprises capecitabine, gemcitabine, nelarabine, or hydroxycarbamide.

49. The method of claim 46, wherein the platinum-based agent comprises carboplatin, oxaliplatin, or cisplatin.

50. The method of claim 46, wherein the topoisomerase I inhibitor comprises camptothecin, irinotecan, or topotecan.

51. The method of claim 46, wherein the topoisomerase II inhibitor comprises etoposide or teniposide.

52. The method of claim 46, wherein the DNA-intercalating agent comprises doxorubicin or liposomal doxorubicin.

53. The method of claim 46, wherein the alkylating agent comprises temozolomide, dactinomycin, melphalan, altretamine, carmustine, bendamustine, busulfan, lomustine, chlorambucil, cyclophosphamide, dacarbazine, altretamine, ifosfamide, or procarbazine.

54. The method of claim 46, wherein the anti -microtubule agent comprises estramustine.

55. The method of claim 46, wherein the taxane-related anti-mitotic agent comprises docetaxel, paclitaxel, cabazitaxel, abraxane, or larotaxel.

56. The method of claim 46, wherein the anti-metabolite comprises 5-fluorouracil, 6- thioguanine, pemetrexed, cytarabine, hydroxyurea, fludarabine, floxuridine, cladribine, pentostatin, or methotrexate.

57. The method of claim 46, wherein the anti-tumor plant alkaloid comprises vinblastine, vincristine, or vinorelbine.

58. The method of claim 46, wherein the anti -estrogen agent comprises tamoxifen, toremifene, raloxifene, lasofoxifene, afimoxifene, arzoxifene, bazedoxifene, fispemifene, ormeloxifene, ospemifene, tesmilifene, trilostane, CHF 4227, fulvestrant, elacestrant, giredestrant, amcenestrant, camizestrant, ataraestane, formestane, exemestane, letrozole, anastrozole, fadrozole; gonadotropin-releasing hormone, leuprolide, leuprolide acetate, enzalutamide, abiraterone acetate, or bicalutamide.

59. The method of claim 46, wherein the protein kinase inhibitor comprises alvocidib, palbociclib, ribociclib, trilaciclib, abemaciclib, crizotinib, NVP-TAE684, ceritinib, alectinib, brigatinib, entrectinib, lorlatinib, vemurafenib, dabrafenib, infigratinib, dovitinib, erdafitinib, BLU-554, AZD4547, trametinib, cobimetinib, binimetinib, selumetinib, ulixertinib, MK-8353, LY-3214966, bevacizumab, axitinib, aflibercept, brivanib, motesanib, pasireotide, sorafenib, erlotinib, linifanib, sunitinib, pazopanib, gefitinib, osimertinib, cetuximab, panitumumab, trastuzumab, neratinib, lapatinib, or cabozantinib.

60. The method of claim 46 or 59, wherein the protein kinase inhibitor comprises a CDK4 / 6 inhibitor.

61. The method of claim 60, wherein the CDK4 / 6 inhibitor comprises palbociclib, ribociclib, trilaciclib, or abemaciclib.

62. The method of claim 46, wherein the phosphatidylinositol 3-kinase inhibitor comprises apelisib, duvelisib, copanlisib, or idelalisib.

63. The method of claim 46, wherein the histone deacetylase inhibitor comprises voninostat.

64. The method of claim 46, wherein the KRAS inhibitor comprises AMG510, MRTX849, JNJ-74699157 / ARS- 3248, Bl 1701963, Bl 1823911, BAY-293, GDC-6036, MRTX1133, or a RAS(ON) inhibitor.

65. The method of claim 46, wherein the immunotherapeutic agent targets at least one of PD1, PD-L1, PD-L2, CTLA-4, LAG3, TIM3, VISTA, BTLA, TIGIT, LAIR1, CD160, 2B4, 4- 1BB, GITR, 0X40, or TGF-beta receptor.

66. The method of claim 65, wherein the immunotherapeutic agent comprises a CTLA-4 inhibitor, a PD1 inhibitor, a PD-L1 inhibitor, a LAG3 inhibitor, or a 4- IBB agonist.

67. The method of claim 66, wherein the CTLA-4 inhibitor comprises tremelimumab or ipilimumab.

68. The method of claim 66, wherein the PD1 inhibitor comprises nivolumab or pembrolizumab.

69. The method of claim 66, wherein the PD-L1 inhibitor comprises atezolizumab, avelumab, or durvalumab.

70. The method of claim 66, wherein the LAG3 inhibitor comprises BMS-986016.

71. The method of claim 46, wherein the epigenetic modulator comprises EZH2a, ARID 1 A, ARID2, PRMT1-9, BRD4, IDH1 / 2, or BCL6.

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